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Title: Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament

Abstract

The bacterial RecA protein plays a role in the complex system of DNA damage repair. Here, we report the functional and structural characterization of the Herbaspirillum seropedicae RecA protein (HsRecA). HsRecA protein is more efficient at displacing SSB protein from ssDNA than Escherichia coli RecA protein. HsRecA also promotes DNA strand exchange more efficiently. The three dimensional structure of HsRecA-ADP/ATP complex has been solved to 1.7 Å resolution. HsRecA protein contains a small N-terminal domain, a central core ATPase domain and a large C-terminal domain, that are similar to homologous bacterial RecA proteins. Comparative structural analysis showed that the N-terminal polymerization motif of archaeal and eukaryotic RecA family proteins are also present in bacterial RecAs. Reconstruction of electrostatic potential from the hexameric structure of HsRecA-ADP/ATP revealed a high positive charge along the inner side, where ssDNA is bound inside the filament. The properties of this surface may explain the greater capacity of HsRecA protein to bind ssDNA, forming a contiguous nucleoprotein filament, displace SSB and promote DNA exchange relative to EcRecA. In conclusion, our functional and structural analyses provide insight into the molecular mechanisms of polymerization of bacterial RecA as a helical nucleoprotein filament.

Authors:
ORCiD logo; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
INCT-FBN; National Institute of General Medical Sciences (NIGMS)
OSTI Identifier:
1347027
DOE Contract Number:  
GM098885
Resource Type:
Journal Article
Resource Relation:
Journal Name: PLoS ONE; Journal Volume: 11; Journal Issue: 7
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Leite, Wellington C., Galvão, Carolina W., Saab, Sérgio C., Iulek, Jorge, Etto, Rafael M., Steffens, Maria B. R., Chitteni-Pattu, Sindhu, Stanage, Tyler, Keck, James L., Cox, Michael M., and Spies, Maria. Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament. United States: N. p., 2016. Web. doi:10.1371/journal.pone.0159871.
Leite, Wellington C., Galvão, Carolina W., Saab, Sérgio C., Iulek, Jorge, Etto, Rafael M., Steffens, Maria B. R., Chitteni-Pattu, Sindhu, Stanage, Tyler, Keck, James L., Cox, Michael M., & Spies, Maria. Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament. United States. doi:10.1371/journal.pone.0159871.
Leite, Wellington C., Galvão, Carolina W., Saab, Sérgio C., Iulek, Jorge, Etto, Rafael M., Steffens, Maria B. R., Chitteni-Pattu, Sindhu, Stanage, Tyler, Keck, James L., Cox, Michael M., and Spies, Maria. Fri . "Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament". United States. doi:10.1371/journal.pone.0159871.
@article{osti_1347027,
title = {Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament},
author = {Leite, Wellington C. and Galvão, Carolina W. and Saab, Sérgio C. and Iulek, Jorge and Etto, Rafael M. and Steffens, Maria B. R. and Chitteni-Pattu, Sindhu and Stanage, Tyler and Keck, James L. and Cox, Michael M. and Spies, Maria},
abstractNote = {The bacterial RecA protein plays a role in the complex system of DNA damage repair. Here, we report the functional and structural characterization of the Herbaspirillum seropedicae RecA protein (HsRecA). HsRecA protein is more efficient at displacing SSB protein from ssDNA than Escherichia coli RecA protein. HsRecA also promotes DNA strand exchange more efficiently. The three dimensional structure of HsRecA-ADP/ATP complex has been solved to 1.7 Å resolution. HsRecA protein contains a small N-terminal domain, a central core ATPase domain and a large C-terminal domain, that are similar to homologous bacterial RecA proteins. Comparative structural analysis showed that the N-terminal polymerization motif of archaeal and eukaryotic RecA family proteins are also present in bacterial RecAs. Reconstruction of electrostatic potential from the hexameric structure of HsRecA-ADP/ATP revealed a high positive charge along the inner side, where ssDNA is bound inside the filament. The properties of this surface may explain the greater capacity of HsRecA protein to bind ssDNA, forming a contiguous nucleoprotein filament, displace SSB and promote DNA exchange relative to EcRecA. In conclusion, our functional and structural analyses provide insight into the molecular mechanisms of polymerization of bacterial RecA as a helical nucleoprotein filament.},
doi = {10.1371/journal.pone.0159871},
journal = {PLoS ONE},
number = 7,
volume = 11,
place = {United States},
year = {Fri Jul 22 00:00:00 EDT 2016},
month = {Fri Jul 22 00:00:00 EDT 2016}
}