Structural characterization of the late competence protein ComFB from Bacillus subtilis

Many bacteria take up DNA from their environment as part of the process of natural transformation. DNA uptake allows microorganisms to gain genetic diversity and can lead to the spread of antibiotic resistance or virulence genes within a microbial population. Development of genetic competence (Com)...

Full description

Bibliographic Details
Main Authors:	Sysoeva, Tatyana A., Bane, Lukas B., Xiao, Daphne Y., Bose, Baundauna, Chilton, Scott S., Gaudet, Rachelle, Burton, Briana M.
Format:	Online
Language:	English
Published:	Portland Press Ltd. 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4381287/

id	pubmed-4381287
recordtype	oai_dc
spelling	pubmed-43812872015-04-07 Structural characterization of the late competence protein ComFB from Bacillus subtilis Sysoeva, Tatyana A. Bane, Lukas B. Xiao, Daphne Y. Bose, Baundauna Chilton, Scott S. Gaudet, Rachelle Burton, Briana M. Original Paper Many bacteria take up DNA from their environment as part of the process of natural transformation. DNA uptake allows microorganisms to gain genetic diversity and can lead to the spread of antibiotic resistance or virulence genes within a microbial population. Development of genetic competence (Com) in Bacillus subtilis is a highly regulated process that culminates in expression of several late competence genes and formation of the DNA uptake apparatus. The late competence operon comF encodes a small protein of unknown function, ComFB. To gain insight into the function of ComFB, we determined its 3D structure via X-ray crystallography. ComFB is a dimer and each subunit consists of four α-helices connected by short loops and one extended β-strand-like stretch. Each subunit contains one zinc-binding site formed by four cysteines, which are unusually spaced in the primary sequence. Using structure- and bioinformatics-guided substitutions we analyzed the inter-subunit interface of the ComFB dimer. Based on these analyses, we conclude that ComFB is an obligate dimer. We also characterized ComFB in vivo and found that this protein is produced in competent cells and is localized to the cytosol. Consistent with previous reports, we showed that deletion of ComFB does not affect DNA uptake function. Combining our results, we conclude that ComFB is unlikely to be a part of the DNA uptake machinery under tested conditions and instead may have a regulatory function. Portland Press Ltd. 2015-03-31 /pmc/articles/PMC4381287/ /pubmed/25423369 http://dx.doi.org/10.1042/BSR20140174 Text en © 2015 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Sysoeva, Tatyana A. Bane, Lukas B. Xiao, Daphne Y. Bose, Baundauna Chilton, Scott S. Gaudet, Rachelle Burton, Briana M.
spellingShingle	Sysoeva, Tatyana A. Bane, Lukas B. Xiao, Daphne Y. Bose, Baundauna Chilton, Scott S. Gaudet, Rachelle Burton, Briana M. Structural characterization of the late competence protein ComFB from Bacillus subtilis
author_facet	Sysoeva, Tatyana A. Bane, Lukas B. Xiao, Daphne Y. Bose, Baundauna Chilton, Scott S. Gaudet, Rachelle Burton, Briana M.
author_sort	Sysoeva, Tatyana A.
title	Structural characterization of the late competence protein ComFB from Bacillus subtilis
title_short	Structural characterization of the late competence protein ComFB from Bacillus subtilis
title_full	Structural characterization of the late competence protein ComFB from Bacillus subtilis
title_fullStr	Structural characterization of the late competence protein ComFB from Bacillus subtilis
title_full_unstemmed	Structural characterization of the late competence protein ComFB from Bacillus subtilis
title_sort	structural characterization of the late competence protein comfb from bacillus subtilis
description	Many bacteria take up DNA from their environment as part of the process of natural transformation. DNA uptake allows microorganisms to gain genetic diversity and can lead to the spread of antibiotic resistance or virulence genes within a microbial population. Development of genetic competence (Com) in Bacillus subtilis is a highly regulated process that culminates in expression of several late competence genes and formation of the DNA uptake apparatus. The late competence operon comF encodes a small protein of unknown function, ComFB. To gain insight into the function of ComFB, we determined its 3D structure via X-ray crystallography. ComFB is a dimer and each subunit consists of four α-helices connected by short loops and one extended β-strand-like stretch. Each subunit contains one zinc-binding site formed by four cysteines, which are unusually spaced in the primary sequence. Using structure- and bioinformatics-guided substitutions we analyzed the inter-subunit interface of the ComFB dimer. Based on these analyses, we conclude that ComFB is an obligate dimer. We also characterized ComFB in vivo and found that this protein is produced in competent cells and is localized to the cytosol. Consistent with previous reports, we showed that deletion of ComFB does not affect DNA uptake function. Combining our results, we conclude that ComFB is unlikely to be a part of the DNA uptake machinery under tested conditions and instead may have a regulatory function.
publisher	Portland Press Ltd.
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4381287/
_version_	1613206134631432192

Structural characterization of the late competence protein ComFB from Bacillus subtilis

Similar Items