SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo

Abstract Genome organization is important for DNA replication, gene expression, and chromosome segregation. In bacteria, two large families of proteins, nucleoid-associated proteins (NAPs) and SMC complexes, play important roles in organizing the genome. NAPs are highly abundant DNA-binding proteins...

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Main Authors: Zhongqing Ren, Lindsey E. Way, Xindan Wang
Format: Article
Language:English
Published: Nature Portfolio 2025-01-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-86946-4
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author Zhongqing Ren
Lindsey E. Way
Xindan Wang
author_facet Zhongqing Ren
Lindsey E. Way
Xindan Wang
author_sort Zhongqing Ren
collection DOAJ
description Abstract Genome organization is important for DNA replication, gene expression, and chromosome segregation. In bacteria, two large families of proteins, nucleoid-associated proteins (NAPs) and SMC complexes, play important roles in organizing the genome. NAPs are highly abundant DNA-binding proteins that can bend, wrap, bridge, and compact DNA, while SMC complexes load onto the chromosome, translocate on the DNA, and extrude DNA loops. Although SMC complexes are capable of traversing the entire chromosome bound by various NAPs in vivo, it is unclear whether SMC translocation is influenced by NAPs. In this study, using Bacillus subtilis as a model system, we expressed a collection of representative bacterial and archaeal DNA-binding proteins that introduce distinct DNA structures and potentially pose different challenges for SMC movement. By fluorescence microscopy and chromatin immunoprecipitation, we observed that these proteins bound to the genome in characteristic manners. Using genome-wide chromosome conformation capture (Hi-C) assays, we found that the SMC complex traversed these DNA-binding proteins without slowing down. Our findings revealed that the DNA-loop-extruding activity of the SMC complex is unaffected by exogenously expressed DNA-binding proteins, which highlights the robustness of SMC motors on the busy chromatin.
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spelling doaj-art-c509eba4f1c24696a1a396a074eef5872025-01-26T12:26:12ZengNature PortfolioScientific Reports2045-23222025-01-0115111310.1038/s41598-025-86946-4SMC translocation is unaffected by an excess of nucleoid associated proteins in vivoZhongqing Ren0Lindsey E. Way1Xindan Wang2Department of Biology, Indiana UniversityDepartment of Biology, Indiana UniversityDepartment of Biology, Indiana UniversityAbstract Genome organization is important for DNA replication, gene expression, and chromosome segregation. In bacteria, two large families of proteins, nucleoid-associated proteins (NAPs) and SMC complexes, play important roles in organizing the genome. NAPs are highly abundant DNA-binding proteins that can bend, wrap, bridge, and compact DNA, while SMC complexes load onto the chromosome, translocate on the DNA, and extrude DNA loops. Although SMC complexes are capable of traversing the entire chromosome bound by various NAPs in vivo, it is unclear whether SMC translocation is influenced by NAPs. In this study, using Bacillus subtilis as a model system, we expressed a collection of representative bacterial and archaeal DNA-binding proteins that introduce distinct DNA structures and potentially pose different challenges for SMC movement. By fluorescence microscopy and chromatin immunoprecipitation, we observed that these proteins bound to the genome in characteristic manners. Using genome-wide chromosome conformation capture (Hi-C) assays, we found that the SMC complex traversed these DNA-binding proteins without slowing down. Our findings revealed that the DNA-loop-extruding activity of the SMC complex is unaffected by exogenously expressed DNA-binding proteins, which highlights the robustness of SMC motors on the busy chromatin.https://doi.org/10.1038/s41598-025-86946-4SMCNAPNucleoid-associated proteinsHUHBsuH-NS
spellingShingle Zhongqing Ren
Lindsey E. Way
Xindan Wang
SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
Scientific Reports
SMC
NAP
Nucleoid-associated proteins
HU
HBsu
H-NS
title SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
title_full SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
title_fullStr SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
title_full_unstemmed SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
title_short SMC translocation is unaffected by an excess of nucleoid associated proteins in vivo
title_sort smc translocation is unaffected by an excess of nucleoid associated proteins in vivo
topic SMC
NAP
Nucleoid-associated proteins
HU
HBsu
H-NS
url https://doi.org/10.1038/s41598-025-86946-4
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AT lindseyeway smctranslocationisunaffectedbyanexcessofnucleoidassociatedproteinsinvivo
AT xindanwang smctranslocationisunaffectedbyanexcessofnucleoidassociatedproteinsinvivo