Cargando…

Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL

Mismatch Repair System corrects mutations arising from DNA replication that escape from DNA polymerase proofreading activity. This system consists of three main proteins, MutS-L-H, responsible for lesion recognition and repair. MutL is a member of GHKL ATPase family and its ATPase cycle has been pro...

Descripción completa

Detalles Bibliográficos
Autores principales: Miguel, Virginia, Correa, Elisa M. E., De Tullio, Luisina, Barra, José L., Argaraña, Carlos E., Villarreal, Marcos A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3724809/
https://www.ncbi.nlm.nih.gov/pubmed/23922851
http://dx.doi.org/10.1371/journal.pone.0069907
_version_ 1782476726466510848
author Miguel, Virginia
Correa, Elisa M. E.
De Tullio, Luisina
Barra, José L.
Argaraña, Carlos E.
Villarreal, Marcos A.
author_facet Miguel, Virginia
Correa, Elisa M. E.
De Tullio, Luisina
Barra, José L.
Argaraña, Carlos E.
Villarreal, Marcos A.
author_sort Miguel, Virginia
collection PubMed
description Mismatch Repair System corrects mutations arising from DNA replication that escape from DNA polymerase proofreading activity. This system consists of three main proteins, MutS-L-H, responsible for lesion recognition and repair. MutL is a member of GHKL ATPase family and its ATPase cycle has been proposed to modulate MutL activity during the repair process. Pseudomonas aeruginosa MutL (PaMutL) contains an N-terminal (NTD) ATPase domain connected by a linker to a C-terminal (CTD) dimerization domain that possesses metal ion-dependent endonuclease activity. With the aim to identify characteristics that allow the PaMutL NTD allosteric control of CTD endonuclease activity, we used an in silico and experimental approach to determine the interaction surfaces of P. aeruginosa NTD (PaNTD), and compared it with the well characterized Escherichia coli MutL NTD (EcNTD). Molecular dynamics simulations of PaNTD and EcNTD bound to or free of adenosine nucleotides showed that a significant difference exists between the behavior of the EcNTD and PaNTD dimerization interface, particularly in the ATP lid. Structure based simulations of MutL homologues with endonuclease activity were performed that allowed an insight of the dimerization interface behavior in this family of proteins. Our experimental results show that, unlike EcNTD, PaNTD is dimeric in presence of ADP. Simulations in mixed solvent allowed us to identify the PaNTD putative DNA binding patch and a putative interaction patch located opposite to the dimerization face. Structure based simulations of PaNTD dimer in presence of ADP or ATP suggest that nucleotide binding could differentially modulate PaNTD protein-protein interactions. Far western assays performed in presence of ADP or ATP are in agreement with our in silico analysis.
format Online
Article
Text
id pubmed-3724809
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37248092013-08-06 Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL Miguel, Virginia Correa, Elisa M. E. De Tullio, Luisina Barra, José L. Argaraña, Carlos E. Villarreal, Marcos A. PLoS One Research Article Mismatch Repair System corrects mutations arising from DNA replication that escape from DNA polymerase proofreading activity. This system consists of three main proteins, MutS-L-H, responsible for lesion recognition and repair. MutL is a member of GHKL ATPase family and its ATPase cycle has been proposed to modulate MutL activity during the repair process. Pseudomonas aeruginosa MutL (PaMutL) contains an N-terminal (NTD) ATPase domain connected by a linker to a C-terminal (CTD) dimerization domain that possesses metal ion-dependent endonuclease activity. With the aim to identify characteristics that allow the PaMutL NTD allosteric control of CTD endonuclease activity, we used an in silico and experimental approach to determine the interaction surfaces of P. aeruginosa NTD (PaNTD), and compared it with the well characterized Escherichia coli MutL NTD (EcNTD). Molecular dynamics simulations of PaNTD and EcNTD bound to or free of adenosine nucleotides showed that a significant difference exists between the behavior of the EcNTD and PaNTD dimerization interface, particularly in the ATP lid. Structure based simulations of MutL homologues with endonuclease activity were performed that allowed an insight of the dimerization interface behavior in this family of proteins. Our experimental results show that, unlike EcNTD, PaNTD is dimeric in presence of ADP. Simulations in mixed solvent allowed us to identify the PaNTD putative DNA binding patch and a putative interaction patch located opposite to the dimerization face. Structure based simulations of PaNTD dimer in presence of ADP or ATP suggest that nucleotide binding could differentially modulate PaNTD protein-protein interactions. Far western assays performed in presence of ADP or ATP are in agreement with our in silico analysis. Public Library of Science 2013-07-26 /pmc/articles/PMC3724809/ /pubmed/23922851 http://dx.doi.org/10.1371/journal.pone.0069907 Text en © 2013 Miguel et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Miguel, Virginia
Correa, Elisa M. E.
De Tullio, Luisina
Barra, José L.
Argaraña, Carlos E.
Villarreal, Marcos A.
Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title_full Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title_fullStr Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title_full_unstemmed Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title_short Analysis of the Interaction Interfaces of the N-Terminal Domain from Pseudomonas aeruginosa MutL
title_sort analysis of the interaction interfaces of the n-terminal domain from pseudomonas aeruginosa mutl
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3724809/
https://www.ncbi.nlm.nih.gov/pubmed/23922851
http://dx.doi.org/10.1371/journal.pone.0069907
work_keys_str_mv AT miguelvirginia analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl
AT correaelisame analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl
AT detullioluisina analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl
AT barrajosel analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl
AT argaranacarlose analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl
AT villarrealmarcosa analysisoftheinteractioninterfacesofthenterminaldomainfrompseudomonasaeruginosamutl