Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining

Kaminski, Andrea M.; Chiruvella, Kishore K.; Ramsden, Dale A.; Bebenek, Katarzyna; Kunkel, Thomas A.; Pedersen, Lars C.

doi:10.1038/s41467-022-31278-4

Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining

Journal Article · Fri Jul 01 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-31278-4· OSTI ID:1879698

^[1]; Chiruvella, Kishore K. ^[2]; ^[2]; ^[1]; ^[1]; ^[1]

National Institutes of Health (NIH), Research Triangle Park, NC (United States). National Institute of Environmental Health Sciences (NIEHS)
Univ. of North Carolina, Chapel Hill, NC (United States). Lineberger Comprehensive Cancer Center

DNA double-strand breaks (DSBs) threaten genomic stability, since their persistence can lead to loss of critical genetic information, chromosomal translocations or rearrangements, and cell death. DSBs can be repaired through the nonhomologous end-joining pathway (NHEJ), which processes and ligates DNA ends efficiently to prevent or minimize sequence loss. Polymerase λ (Polλ), one of the Family X polymerases, fills sequence gaps of DSB substrates with a strict specificity for a base-paired primer terminus. There is little information regarding Polλ’s approach to engaging such substrates. We used in vitro polymerization and cell-based NHEJ assays to explore the contributions of conserved loop regions toward DSB substrate specificity and utilization. In addition, we present multiple crystal structures of Polλ in synapsis with varying biologically relevant DSB end configurations, revealing how key structural features and hydrogen bonding networks work in concert to stabilize these tenuous, potentially cytotoxic DNA lesions during NHEJ.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

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

Grant/Contract Number:: W-31109-ENG-38

OSTI ID:: 1879698

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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