ssDNA is an allosteric regulator of the C. crescentus SOS-independent DNA damage response transcription activator, DriD

Gozzi, Kevin; Salinas, Raul; Nguyen, Viet D.; Laub, Michael T.; Schumacher, Maria A.

doi:10.1101/gad.349541.122

Title: ssDNA is an allosteric regulator of the C. crescentus SOS-independent DNA damage response transcription activator, DriD

Journal Article · 26 May 2022 · Genes & Development

DOI: https://doi.org/10.1101/gad.349541.122 · OSTI ID:1903864

Gozzi, Kevin ^[1]; Salinas, Raul ^[2]; Nguyen, Viet D. ^[2]; Laub, Michael T. ^[3];

^[2]

Massachusetts Institute of Technology (MIT), Cambridge, MA (United States); OSTI
Duke University, Durham, NC (United States). Medical Center
Massachusetts Institute of Technology (MIT), Cambridge, MA (United States)

DNA damage repair systems are critical for genomic integrity. However, they must be coordinated with DNA replication and cell division to ensure accurate genomic transmission. In most bacteria, this coordination is mediated by the SOS response through LexA, which triggers a halt in cell division until repair is completed. Recently, an SOS-independent damage response system was revealed in Caulobacter crescentus. This pathway is controlled by the transcription activator, DriD, but how DriD senses and signals DNA damage is unknown. To address this question, we performed biochemical, cellular, and structural studies. We show that DriD binds a specific promoter DNA site via its N-terminal HTH domain to activate transcription of genes, including the cell division inhibitor didA. A structure of the C-terminal portion of DriD revealed a WYL motif domain linked to a WCX dimerization domain. Strikingly, we found that DriD binds ssDNA between the WYL and WCX domains. Comparison of apo and ssDNA-bound DriD structures reveals that ssDNA binding orders and orients the DriD domains, indicating a mechanism for ssDNA-mediated operator DNA binding activation. Biochemical and in vivo studies support the structural model. Our data thus reveal the molecular mechanism underpinning an SOS-independent DNA damage repair pathway.

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Research Organization:: Massachusetts Institute of Technology (MIT), Cambridge, MA (United States); Duke University, Durham, NC (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1903864

Journal Information:: Genes & Development, Journal Name: Genes & Development Journal Issue: 9-10 Vol. 36; ISSN 0890-9369

Publisher:: Cold Springs Harbor Laboratory PressCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Title: ssDNA is an allosteric regulator of the C. crescentus SOS-independent DNA damage response transcription activator, DriD

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