SlideShare una empresa de Scribd logo
1 de 1
Descargar para leer sin conexión
STRUCTURAL	
  ORDER	
  AND	
  DISORDER	
  DICTATE	
  SEQUENCE	
  AND	
  FUNCTIONAL	
  
                                                 EVOLUTION	
  OF	
  THE	
  PAPILLOMAVIRUS	
  E7	
  PROTEIN	
  
                                                    LUCIA	
  B.	
  CHEMES¶,	
  JULIANA	
  GLAVINA§,	
  CRISTINA	
  MARINO-­‐BUSLJE¶,	
  GONZALO	
  DE	
  PRAT-­‐GAY¶	
  	
  
                                                                                             AND	
  IGNACIO	
  E.	
  SANCHEZ§	
  
                               ¶PROTEIN	
  STRUCTURE,	
  FUNCTION	
  AND	
  ENGINEERING	
  LABORATORY,	
  FUNDACION	
  INSTITUTO	
  LELOIR	
  AND	
  IIBBA-­‐CONICET,	
  BUENOS	
  AIRES,	
  ARGENTINA.	
  §PROTEIN	
  

                              PHYSIOLOGY	
  LABORATORY,	
  DEPARTAMENTO	
  DE	
  QUIMICA	
  BIOLOGICA,	
  FACULTAD	
  DE	
  CIENCIAS	
  EXACTAS	
  Y	
  NATURALES-­‐UNIVERSIDAD	
  DE	
  BUENOS	
  AIRES,	
  ARGENTINA	
  



 INTRODUCTION	
  
 E7	
  is	
  the	
  main	
  transforming	
  protein	
  in	
  papillomaviruses	
  and	
  plays	
  an	
  important	
  role	
  in	
  oncogenesis	
  [1].	
  The	
  globular	
  C-­‐terminal	
  
 domain	
   (E7C)	
   mediates	
   zinc	
   binding	
   and	
   homodimeriza+on	
   [2].	
   The	
   intrinsically	
   disordered	
   N-­‐terminal	
   domain	
   (E7N)	
   harbors	
  
 several	
   linear	
   mo+fs	
   that	
   mediate	
   interac+on	
   with	
   cellular	
   targets,	
   including	
   the	
   high	
   affinity	
   LxCxE	
   binding	
   site	
   for	
   the	
  
 Re+noblastoma	
  protein	
  (Rb)	
  [3].	
  We	
  have	
  analyzed	
  sequence	
  and	
  func+onal	
  evolu+on	
  of	
  E7	
  using	
  210	
  natural	
  sequences.	
  	
  


                                          E7N DOMAIN CONSERVATION!                                                                                                                E7C DOMAIN CONSERVATION!
             RbAB (E2F SITE)                             RbAB (LxCxE SITE)                                   RbAB (LxCxE SITE)
               p600 p300                                                                                                                                              RbC, p21CIP, p27KIP, TBP, AP1, Mi2B, IGFBP3, S4 proteasome, MPP2
                                                           P107 p130 p21CIP                                     TBP F-Actin
           IRF-1 Cullin-2 FHL2                             p300 IRF-1 FHL2                                      CKII HPV-E2C                                               IRF-1, h-TID, pCAF, Cullin2-UBC, E2F1, FHL2, NuMA, DNMT1

                  CR1-Helix!                                     LxCxE!                                           CKII-PEST!                                                                                                         NES!




  * Coevolving residue pairs"                                  *         **
                                                                          *                                        *                                      A
                                                                                                                                                                              *                        *           *
                                                                                                                                                                                                                   *           B
                                                                                                                                                                                                                                            *
                                                                                                                                                                                    CONSERVATION!                                    CO-EVOLUTION!
   A                 INFORMATION CONTENT!                                B                      CKII-PEST REGION!




                                                                                                                                                                          Zn-binding cysteines"            Surface residues"                    D61/T72"
                                                                                                                                                                          Monomer interface"               Dimer interface"                     C45/Q56"
The sequence logos [4] show that E7N is as conserved as E7C in spite of the lack of a stable structure (figure A). The
highly conserved E7N motifs are separated by variable regions. The CR1 region shows high conservation at the                                         The sequence logo [4] for the C-terminal domain displays (1) four cysteine residues involved in zinc binding (positions
helix-forming Rb-targeting residues 6-13 and at uncharacterized residues 1-3. The LxCxE motif also shows                                             44, 47, 77 and 80, displayed in red in figure A), (2) a highly conserved leucine-rich region that acts as a nuclear
conservation at residues that are outside the canonical motif (pos. 19, 23, 25 and 26). One third of the E7 sequences                                export signal, (3) six conserved positions (displayed in blue in figure A) that form the core of each monomer, and (4)
lack a CKII phosphorylation site, while only 2.5% of them lack a stretch of acidic residues (n>3) (figure B). The tight                              six conserved positions (displayed in cyan in figure A) that stabilize the dimer interface. Four conserved residues are
restriction in sequence separation between the LxCxE motif and the CKII/PEST region together with the coevolution                                    surface exposed (yellow in figure A). A mutual information analysis [5] reveals two pairs of coevolving amino acid
of residues 25 and 29 [5] (black asterisks), suggests that the two motifs form an evolutionary and functional unit.                                  positions that form contacts across the dimerization interface (figure B).



                                 EVOLUTION OF E7N LINEAR MOTIFS!                                                                                                                       E7C CYSTEINE CLUSTERS!




                                                                                                                                                                              REGION 2"            Zn-binding cysteines"
                                                                                                                                                                              REGION 1"            Zinc ion"

                                                                                                                                                     Two E7C sequence regions show high frequencies of Cysteine (6 to 21%), with most E7 proteins having at least one
                                                                                                                                                     extra cysteine in addition to the two canonical CxxC motifs. One cysteine-rich region (blue) is close in space and
                                                                                                                                                     sequence to the first CxxC motif and the zinc ion. The second region (green) is close in space to the zinc ion
                                                                                                                                                     coordinated by the opposite monomer. The additional cysteines may stabilize alternative conformations of the domain
                                                                                                                                                     through non-native coordination of the zinc ion.




                                                                                                                                                                         CONSERVED PEPTIDE-BINDING SITE IN E7C!
                                                                                                                                                                            HPV45 E7C (PDB 2B9D)                                   PYGO1_MOUSE (PDB 2YYR)
                                                                                                                                                           P21-binding site                       Information content                       H3-binding site
The phylogenetic analysis suggests that the LxCxE motif, the acidic stretch and the CKII sites have changed several
times during papillomavirus evolution. Changes in sequence of the LxCxE motif are coupled to changes in phenotype
(delta, gamma and alpha 2 sp.), pointing to adaptive evolution events. In reptilian, avian and some artiodactyl
papillomaviruses, E7N is substituted by a domain with no sequence similarity to canonical E7N sequences.
Whenever the LxCxE motif is present, the acidic stretch follows, further supporting the functional association between
them. Gamma papillomaviruses often harbor an LxSxE motif. (Figure adapted from [6])



 CONCLUDING	
  REMARKS	
  
 • 	
   	
  Sequence	
  evolu+on	
  in	
  the	
  disordered	
  E7N	
  domain	
  shows	
  that	
  some	
  of	
  its	
  short	
  
 func+onal	
   mo+fs	
   evolve	
   in	
   a	
   coordinate	
   manner	
   and	
   that	
   the	
   domain	
   has	
   been	
  
 subject	
   to	
   several	
   episodes	
   of	
   adap+ve	
   evolu+on.	
   The	
   high	
   func+onal	
   density	
   within	
                                  PUTATIVE E7C BINDING MOTIF                                      E7C/PYGO1
 E7N	
  could	
  explain	
  the	
  large	
  number	
  of	
  targets	
  found	
  for	
  this	
  small	
  protein.	
  
 • 	
  	
  	
  	
  Evolu+on	
  of	
  the	
  E7C	
  domain	
  is	
  dictated	
  by	
  dimeriza+on,	
  canonical	
  zinc	
  binding	
  by	
  
 the	
  two	
  CxxC	
  mo+fs	
  and	
  likely	
  also	
  by	
  zinc	
  binding	
  by	
  unpaired	
  cysteines	
  and	
  binding	
  
 of	
  short	
  Ser/Pro-­‐rich	
  sequences	
  within	
  host	
  proteins.	
  
                                                                                                                                                     HPV45 E7C binds to a short peptide from the host cellular protein p21 [2]. NMR measurements indicate that the
REFERENCES	
                                                                                                                                         interaction is mediated by certain residues from each monomer's exposed surface (upper panel, left). The proposed
[1] Chemes LB et al. Intrinsic disorder in the human papollomavirus E7 protein. In: Flexible Viruses, Uversky DN and Longhi S. Eds. In press.
[2] Ohlenschlager O et al. Solution sturcture of the partially folded high-risk human papilloma virus 45 oncoprotein E7. Oncogene 2006, 25:5953-9.
                                                                                                                                                     site overlaps well with a conserved surface patch in E7C (upper panel, middle), suggesting that peptide binding is a
[3] Lee JO et al. Structure of the retinoblastoma tumour suppressor pocket domain bound to a peptide from HPV E7. Nature 1998, 361:859-65.           property shared by most E7C domains.
[4] Schneider TD, Stephens RM. Sequence logos: a new way to display consensus sequences. Nucleic Acids Res. 1990, 18:6097-100.                       The structures of the E7C domain and host phd domains superimpose well [7] (lower panel, right), pointing to a
[5] Marino Buslje C et al. Correction for phylogeny, small number of observations and data redundancy improves the identification of coevolving
amino acid pairs using mutual information. Bioinformatics 2009, 25:1125-1131
                                                                                                                                                     plausible evolutionary origin for E7C. Many phd domains bind peptides at a site that corresponds to the proposed
[6] Bravo IG et al. The clinical importance of understanding the evolution of papillomaviruses. Trends in Microbiology 2010, 18:432-438.             functional surface of E7C (upper panel, right). A motif search in the sequences of E7C targets [8] suggests that the
[7] Suhrer S et al.. COPS-a novel workbench for explorations in fold space. Nucleic Acids Res. 2009, 37(Web Server issue):W539-544.                  domain binds peptides rich in proline and serine residues (lower panel, left).
[8] Radusky L et al. Discovery of functional protein linear motifs using a greedy algorithm and information theory. POSTER.

Más contenido relacionado

Destacado

Cutting The Trees Of Knowledge
Cutting The Trees Of KnowledgeCutting The Trees Of Knowledge
Cutting The Trees Of KnowledgeAnnie Chiv
 
09 Nick Stanhope
09 Nick Stanhope09 Nick Stanhope
09 Nick StanhopeABMcentrum
 
01 andrew payne
01 andrew payne01 andrew payne
01 andrew payneABMcentrum
 
Comunicacion interna carlos iii
Comunicacion interna carlos iiiComunicacion interna carlos iii
Comunicacion interna carlos iiiAdrián Segovia
 
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...museum-digital
 

Destacado (15)

About using new descriptors for cheminformatics
About using new descriptors for cheminformaticsAbout using new descriptors for cheminformatics
About using new descriptors for cheminformatics
 
Cooperatividad en la Expresión Génica: Abordaje Estocástico
Cooperatividad en la Expresión Génica: Abordaje EstocásticoCooperatividad en la Expresión Génica: Abordaje Estocástico
Cooperatividad en la Expresión Génica: Abordaje Estocástico
 
Signals of Evolution: Conservation, Specificity Determining Positions and Coe...
Signals of Evolution: Conservation, Specificity Determining Positions and Coe...Signals of Evolution: Conservation, Specificity Determining Positions and Coe...
Signals of Evolution: Conservation, Specificity Determining Positions and Coe...
 
Cutting The Trees Of Knowledge
Cutting The Trees Of KnowledgeCutting The Trees Of Knowledge
Cutting The Trees Of Knowledge
 
09 Nick Stanhope
09 Nick Stanhope09 Nick Stanhope
09 Nick Stanhope
 
01 andrew payne
01 andrew payne01 andrew payne
01 andrew payne
 
Stuart Lee
Stuart LeeStuart Lee
Stuart Lee
 
Comunicacion interna carlos iii
Comunicacion interna carlos iiiComunicacion interna carlos iii
Comunicacion interna carlos iii
 
Modelado de la proteína p35 de toxoplasma gondii
Modelado de la proteína p35 de toxoplasma gondiiModelado de la proteína p35 de toxoplasma gondii
Modelado de la proteína p35 de toxoplasma gondii
 
Discovery Of Functional Protein Linear Motifs Using a Greaddy Algorithm and I...
Discovery Of Functional Protein Linear Motifs Using a Greaddy Algorithm and I...Discovery Of Functional Protein Linear Motifs Using a Greaddy Algorithm and I...
Discovery Of Functional Protein Linear Motifs Using a Greaddy Algorithm and I...
 
La Unidad de Bioinformática del INTA
La Unidad de Bioinformática del INTALa Unidad de Bioinformática del INTA
La Unidad de Bioinformática del INTA
 
Predicting peptide/MHC interactions: Application to epitope identification an...
Predicting peptide/MHC interactions: Application to epitope identification an...Predicting peptide/MHC interactions: Application to epitope identification an...
Predicting peptide/MHC interactions: Application to epitope identification an...
 
NIF
NIFNIF
NIF
 
Bioinformatica Proteinas
Bioinformatica ProteinasBioinformatica Proteinas
Bioinformatica Proteinas
 
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...
www.museum-digital.de * project presentation at athena-workshop in Stuttgart,...
 

Más de Asociación Argentina de Bioinformática y Biología Computacional

Más de Asociación Argentina de Bioinformática y Biología Computacional (8)

Design of degenerated primers from bioinformatics online software for putativ...
Design of degenerated primers from bioinformatics online software for putativ...Design of degenerated primers from bioinformatics online software for putativ...
Design of degenerated primers from bioinformatics online software for putativ...
 
A structure-function analysis of s HSPs in plants
A structure-function analysis of s HSPs in plantsA structure-function analysis of s HSPs in plants
A structure-function analysis of s HSPs in plants
 
Data balancing for phenotype classification based on SNPs
Data balancing for phenotype classification based on SNPsData balancing for phenotype classification based on SNPs
Data balancing for phenotype classification based on SNPs
 
Gene selection via significant subset using silhouette index
Gene selection via significant subset using silhouette indexGene selection via significant subset using silhouette index
Gene selection via significant subset using silhouette index
 
Bolstered error estimation for discrete classifier applied to genomic signal ...
Bolstered error estimation for discrete classifier applied to genomic signal ...Bolstered error estimation for discrete classifier applied to genomic signal ...
Bolstered error estimation for discrete classifier applied to genomic signal ...
 
Biopython: Overview, State of the Art and Outlook
Biopython: Overview, State of the Art and OutlookBiopython: Overview, State of the Art and Outlook
Biopython: Overview, State of the Art and Outlook
 
¿Cuál es la estabilidad relevante de las proteínas?
¿Cuál es la estabilidad relevante de las proteínas?¿Cuál es la estabilidad relevante de las proteínas?
¿Cuál es la estabilidad relevante de las proteínas?
 
Biogeografía histórica y Análisis de Vicarianza: Una perspectiva computacional
Biogeografía histórica y Análisis de Vicarianza: Una perspectiva computacionalBiogeografía histórica y Análisis de Vicarianza: Una perspectiva computacional
Biogeografía histórica y Análisis de Vicarianza: Una perspectiva computacional
 

Último

Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024D Cloud Solutions
 
Machine Learning Model Validation (Aijun Zhang 2024).pdf
Machine Learning Model Validation (Aijun Zhang 2024).pdfMachine Learning Model Validation (Aijun Zhang 2024).pdf
Machine Learning Model Validation (Aijun Zhang 2024).pdfAijun Zhang
 
UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7DianaGray10
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxGDSC PJATK
 
NIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopNIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopBachir Benyammi
 
Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?SANGHEE SHIN
 
Basic Building Blocks of Internet of Things.
Basic Building Blocks of Internet of Things.Basic Building Blocks of Internet of Things.
Basic Building Blocks of Internet of Things.YounusS2
 
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...Will Schroeder
 
Introduction to Quantum Computing
Introduction to Quantum ComputingIntroduction to Quantum Computing
Introduction to Quantum ComputingGDSC PJATK
 
Spring24-Release Overview - Wellingtion User Group-1.pdf
Spring24-Release Overview - Wellingtion User Group-1.pdfSpring24-Release Overview - Wellingtion User Group-1.pdf
Spring24-Release Overview - Wellingtion User Group-1.pdfAnna Loughnan Colquhoun
 
Empowering Africa's Next Generation: The AI Leadership Blueprint
Empowering Africa's Next Generation: The AI Leadership BlueprintEmpowering Africa's Next Generation: The AI Leadership Blueprint
Empowering Africa's Next Generation: The AI Leadership BlueprintMahmoud Rabie
 
UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8DianaGray10
 
Computer 10: Lesson 10 - Online Crimes and Hazards
Computer 10: Lesson 10 - Online Crimes and HazardsComputer 10: Lesson 10 - Online Crimes and Hazards
Computer 10: Lesson 10 - Online Crimes and HazardsSeth Reyes
 
Babel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxBabel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxYounusS2
 
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationUsing IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationIES VE
 
UiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPathCommunity
 
AI Fame Rush Review – Virtual Influencer Creation In Just Minutes
AI Fame Rush Review – Virtual Influencer Creation In Just MinutesAI Fame Rush Review – Virtual Influencer Creation In Just Minutes
AI Fame Rush Review – Virtual Influencer Creation In Just MinutesMd Hossain Ali
 
Digital magic. A small project for controlling smart light bulbs.
Digital magic. A small project for controlling smart light bulbs.Digital magic. A small project for controlling smart light bulbs.
Digital magic. A small project for controlling smart light bulbs.francesco barbera
 
Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024SkyPlanner
 
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...DianaGray10
 

Último (20)

Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024
 
Machine Learning Model Validation (Aijun Zhang 2024).pdf
Machine Learning Model Validation (Aijun Zhang 2024).pdfMachine Learning Model Validation (Aijun Zhang 2024).pdf
Machine Learning Model Validation (Aijun Zhang 2024).pdf
 
UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptx
 
NIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopNIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 Workshop
 
Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?
 
Basic Building Blocks of Internet of Things.
Basic Building Blocks of Internet of Things.Basic Building Blocks of Internet of Things.
Basic Building Blocks of Internet of Things.
 
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...
Apres-Cyber - The Data Dilemma: Bridging Offensive Operations and Machine Lea...
 
Introduction to Quantum Computing
Introduction to Quantum ComputingIntroduction to Quantum Computing
Introduction to Quantum Computing
 
Spring24-Release Overview - Wellingtion User Group-1.pdf
Spring24-Release Overview - Wellingtion User Group-1.pdfSpring24-Release Overview - Wellingtion User Group-1.pdf
Spring24-Release Overview - Wellingtion User Group-1.pdf
 
Empowering Africa's Next Generation: The AI Leadership Blueprint
Empowering Africa's Next Generation: The AI Leadership BlueprintEmpowering Africa's Next Generation: The AI Leadership Blueprint
Empowering Africa's Next Generation: The AI Leadership Blueprint
 
UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8
 
Computer 10: Lesson 10 - Online Crimes and Hazards
Computer 10: Lesson 10 - Online Crimes and HazardsComputer 10: Lesson 10 - Online Crimes and Hazards
Computer 10: Lesson 10 - Online Crimes and Hazards
 
Babel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxBabel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptx
 
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationUsing IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
 
UiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation Developers
 
AI Fame Rush Review – Virtual Influencer Creation In Just Minutes
AI Fame Rush Review – Virtual Influencer Creation In Just MinutesAI Fame Rush Review – Virtual Influencer Creation In Just Minutes
AI Fame Rush Review – Virtual Influencer Creation In Just Minutes
 
Digital magic. A small project for controlling smart light bulbs.
Digital magic. A small project for controlling smart light bulbs.Digital magic. A small project for controlling smart light bulbs.
Digital magic. A small project for controlling smart light bulbs.
 
Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024
 
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...
Connector Corner: Extending LLM automation use cases with UiPath GenAI connec...
 

Structural Order and Disorder Dictate Sequence And Functional Evolution of the Papillomavirus E7 Protein

  • 1. STRUCTURAL  ORDER  AND  DISORDER  DICTATE  SEQUENCE  AND  FUNCTIONAL   EVOLUTION  OF  THE  PAPILLOMAVIRUS  E7  PROTEIN   LUCIA  B.  CHEMES¶,  JULIANA  GLAVINA§,  CRISTINA  MARINO-­‐BUSLJE¶,  GONZALO  DE  PRAT-­‐GAY¶     AND  IGNACIO  E.  SANCHEZ§   ¶PROTEIN  STRUCTURE,  FUNCTION  AND  ENGINEERING  LABORATORY,  FUNDACION  INSTITUTO  LELOIR  AND  IIBBA-­‐CONICET,  BUENOS  AIRES,  ARGENTINA.  §PROTEIN   PHYSIOLOGY  LABORATORY,  DEPARTAMENTO  DE  QUIMICA  BIOLOGICA,  FACULTAD  DE  CIENCIAS  EXACTAS  Y  NATURALES-­‐UNIVERSIDAD  DE  BUENOS  AIRES,  ARGENTINA   INTRODUCTION   E7  is  the  main  transforming  protein  in  papillomaviruses  and  plays  an  important  role  in  oncogenesis  [1].  The  globular  C-­‐terminal   domain   (E7C)   mediates   zinc   binding   and   homodimeriza+on   [2].   The   intrinsically   disordered   N-­‐terminal   domain   (E7N)   harbors   several   linear   mo+fs   that   mediate   interac+on   with   cellular   targets,   including   the   high   affinity   LxCxE   binding   site   for   the   Re+noblastoma  protein  (Rb)  [3].  We  have  analyzed  sequence  and  func+onal  evolu+on  of  E7  using  210  natural  sequences.     E7N DOMAIN CONSERVATION! E7C DOMAIN CONSERVATION! RbAB (E2F SITE) RbAB (LxCxE SITE) RbAB (LxCxE SITE) p600 p300 RbC, p21CIP, p27KIP, TBP, AP1, Mi2B, IGFBP3, S4 proteasome, MPP2 P107 p130 p21CIP TBP F-Actin IRF-1 Cullin-2 FHL2 p300 IRF-1 FHL2 CKII HPV-E2C IRF-1, h-TID, pCAF, Cullin2-UBC, E2F1, FHL2, NuMA, DNMT1 CR1-Helix! LxCxE! CKII-PEST! NES! * Coevolving residue pairs" * ** * * A * * * * B * CONSERVATION! CO-EVOLUTION! A INFORMATION CONTENT! B CKII-PEST REGION! Zn-binding cysteines" Surface residues" D61/T72" Monomer interface" Dimer interface" C45/Q56" The sequence logos [4] show that E7N is as conserved as E7C in spite of the lack of a stable structure (figure A). The highly conserved E7N motifs are separated by variable regions. The CR1 region shows high conservation at the The sequence logo [4] for the C-terminal domain displays (1) four cysteine residues involved in zinc binding (positions helix-forming Rb-targeting residues 6-13 and at uncharacterized residues 1-3. The LxCxE motif also shows 44, 47, 77 and 80, displayed in red in figure A), (2) a highly conserved leucine-rich region that acts as a nuclear conservation at residues that are outside the canonical motif (pos. 19, 23, 25 and 26). One third of the E7 sequences export signal, (3) six conserved positions (displayed in blue in figure A) that form the core of each monomer, and (4) lack a CKII phosphorylation site, while only 2.5% of them lack a stretch of acidic residues (n>3) (figure B). The tight six conserved positions (displayed in cyan in figure A) that stabilize the dimer interface. Four conserved residues are restriction in sequence separation between the LxCxE motif and the CKII/PEST region together with the coevolution surface exposed (yellow in figure A). A mutual information analysis [5] reveals two pairs of coevolving amino acid of residues 25 and 29 [5] (black asterisks), suggests that the two motifs form an evolutionary and functional unit. positions that form contacts across the dimerization interface (figure B). EVOLUTION OF E7N LINEAR MOTIFS! E7C CYSTEINE CLUSTERS! REGION 2" Zn-binding cysteines" REGION 1" Zinc ion" Two E7C sequence regions show high frequencies of Cysteine (6 to 21%), with most E7 proteins having at least one extra cysteine in addition to the two canonical CxxC motifs. One cysteine-rich region (blue) is close in space and sequence to the first CxxC motif and the zinc ion. The second region (green) is close in space to the zinc ion coordinated by the opposite monomer. The additional cysteines may stabilize alternative conformations of the domain through non-native coordination of the zinc ion. CONSERVED PEPTIDE-BINDING SITE IN E7C! HPV45 E7C (PDB 2B9D) PYGO1_MOUSE (PDB 2YYR) P21-binding site Information content H3-binding site The phylogenetic analysis suggests that the LxCxE motif, the acidic stretch and the CKII sites have changed several times during papillomavirus evolution. Changes in sequence of the LxCxE motif are coupled to changes in phenotype (delta, gamma and alpha 2 sp.), pointing to adaptive evolution events. In reptilian, avian and some artiodactyl papillomaviruses, E7N is substituted by a domain with no sequence similarity to canonical E7N sequences. Whenever the LxCxE motif is present, the acidic stretch follows, further supporting the functional association between them. Gamma papillomaviruses often harbor an LxSxE motif. (Figure adapted from [6]) CONCLUDING  REMARKS   •     Sequence  evolu+on  in  the  disordered  E7N  domain  shows  that  some  of  its  short   func+onal   mo+fs   evolve   in   a   coordinate   manner   and   that   the   domain   has   been   subject   to   several   episodes   of   adap+ve   evolu+on.   The   high   func+onal   density   within   PUTATIVE E7C BINDING MOTIF E7C/PYGO1 E7N  could  explain  the  large  number  of  targets  found  for  this  small  protein.   •         Evolu+on  of  the  E7C  domain  is  dictated  by  dimeriza+on,  canonical  zinc  binding  by   the  two  CxxC  mo+fs  and  likely  also  by  zinc  binding  by  unpaired  cysteines  and  binding   of  short  Ser/Pro-­‐rich  sequences  within  host  proteins.   HPV45 E7C binds to a short peptide from the host cellular protein p21 [2]. NMR measurements indicate that the REFERENCES   interaction is mediated by certain residues from each monomer's exposed surface (upper panel, left). The proposed [1] Chemes LB et al. Intrinsic disorder in the human papollomavirus E7 protein. In: Flexible Viruses, Uversky DN and Longhi S. Eds. In press. [2] Ohlenschlager O et al. Solution sturcture of the partially folded high-risk human papilloma virus 45 oncoprotein E7. Oncogene 2006, 25:5953-9. site overlaps well with a conserved surface patch in E7C (upper panel, middle), suggesting that peptide binding is a [3] Lee JO et al. Structure of the retinoblastoma tumour suppressor pocket domain bound to a peptide from HPV E7. Nature 1998, 361:859-65. property shared by most E7C domains. [4] Schneider TD, Stephens RM. Sequence logos: a new way to display consensus sequences. Nucleic Acids Res. 1990, 18:6097-100. The structures of the E7C domain and host phd domains superimpose well [7] (lower panel, right), pointing to a [5] Marino Buslje C et al. Correction for phylogeny, small number of observations and data redundancy improves the identification of coevolving amino acid pairs using mutual information. Bioinformatics 2009, 25:1125-1131 plausible evolutionary origin for E7C. Many phd domains bind peptides at a site that corresponds to the proposed [6] Bravo IG et al. The clinical importance of understanding the evolution of papillomaviruses. Trends in Microbiology 2010, 18:432-438. functional surface of E7C (upper panel, right). A motif search in the sequences of E7C targets [8] suggests that the [7] Suhrer S et al.. COPS-a novel workbench for explorations in fold space. Nucleic Acids Res. 2009, 37(Web Server issue):W539-544. domain binds peptides rich in proline and serine residues (lower panel, left). [8] Radusky L et al. Discovery of functional protein linear motifs using a greedy algorithm and information theory. POSTER.