Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro

Leigh A. Smith, Alena V. Makarova, Laura Samson, Katherine E. Thiesen, Alok Dhar, Tadayoshi Bessho

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Repair of DNA interstrand cross-links in mammalian cells involves several biochemically distinctive processes, including the release of one of the cross-linked strands and translesion DNA synthesis (TLS). In this report, we investigated the in vitro TLS activity of a psoralen DNA interstrand cross-link by three DNA repair polymerases, DNA polymerases β, κ, and l. DNA polymerase β is capable of bypassing a psoralen cross-link with a low efficiency. Cell extracts prepared from DNA polymerase β knockout mouse embryonic fibroblasts showed a reduced bypass activity of the psoralen cross-link, and purified DNA polymerase β restored the bypass activity. In addition, DNA polymerase l misincorporated thymine across the psoralen cross-link and DNA polymerase κ extended these mispaired primer ends, suggesting that DNA polymerase l may serve as an inserter and DNA polymerase κ may play a role as an extender in the repair of psoralen DNA interstrand cross-links. The results demonstrated here indicate that multiple DNA polymerases could participate in TLS steps in mammalian DNA interstrand cross-link repair.

Original languageEnglish (US)
Pages (from-to)8931-8938
Number of pages8
JournalBiochemistry
Volume51
Issue number44
DOIs
StatePublished - Nov 6 2012

Fingerprint

Ficusin
DNA-Directed DNA Polymerase
DNA
Repair
DNA Repair
In Vitro Techniques
Thymine
Fibroblasts
Cell Extracts
Knockout Mice
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Smith, L. A., Makarova, A. V., Samson, L., Thiesen, K. E., Dhar, A., & Bessho, T. (2012). Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro. Biochemistry, 51(44), 8931-8938. https://doi.org/10.1021/bi3008565

Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro. / Smith, Leigh A.; Makarova, Alena V.; Samson, Laura; Thiesen, Katherine E.; Dhar, Alok; Bessho, Tadayoshi.

In: Biochemistry, Vol. 51, No. 44, 06.11.2012, p. 8931-8938.

Research output: Contribution to journalArticle

Smith, LA, Makarova, AV, Samson, L, Thiesen, KE, Dhar, A & Bessho, T 2012, 'Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro', Biochemistry, vol. 51, no. 44, pp. 8931-8938. https://doi.org/10.1021/bi3008565
Smith, Leigh A. ; Makarova, Alena V. ; Samson, Laura ; Thiesen, Katherine E. ; Dhar, Alok ; Bessho, Tadayoshi. / Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro. In: Biochemistry. 2012 ; Vol. 51, No. 44. pp. 8931-8938.
@article{cb955e40992a40fd9e07dbcf0d52784d,
title = "Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro",
abstract = "Repair of DNA interstrand cross-links in mammalian cells involves several biochemically distinctive processes, including the release of one of the cross-linked strands and translesion DNA synthesis (TLS). In this report, we investigated the in vitro TLS activity of a psoralen DNA interstrand cross-link by three DNA repair polymerases, DNA polymerases β, κ, and l. DNA polymerase β is capable of bypassing a psoralen cross-link with a low efficiency. Cell extracts prepared from DNA polymerase β knockout mouse embryonic fibroblasts showed a reduced bypass activity of the psoralen cross-link, and purified DNA polymerase β restored the bypass activity. In addition, DNA polymerase l misincorporated thymine across the psoralen cross-link and DNA polymerase κ extended these mispaired primer ends, suggesting that DNA polymerase l may serve as an inserter and DNA polymerase κ may play a role as an extender in the repair of psoralen DNA interstrand cross-links. The results demonstrated here indicate that multiple DNA polymerases could participate in TLS steps in mammalian DNA interstrand cross-link repair.",
author = "Smith, {Leigh A.} and Makarova, {Alena V.} and Laura Samson and Thiesen, {Katherine E.} and Alok Dhar and Tadayoshi Bessho",
year = "2012",
month = "11",
day = "6",
doi = "10.1021/bi3008565",
language = "English (US)",
volume = "51",
pages = "8931--8938",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "44",

}

TY - JOUR

T1 - Bypass of a psoralen DNA interstrand cross-link by DNA polymerases β, l, and κ in Vitro

AU - Smith, Leigh A.

AU - Makarova, Alena V.

AU - Samson, Laura

AU - Thiesen, Katherine E.

AU - Dhar, Alok

AU - Bessho, Tadayoshi

PY - 2012/11/6

Y1 - 2012/11/6

N2 - Repair of DNA interstrand cross-links in mammalian cells involves several biochemically distinctive processes, including the release of one of the cross-linked strands and translesion DNA synthesis (TLS). In this report, we investigated the in vitro TLS activity of a psoralen DNA interstrand cross-link by three DNA repair polymerases, DNA polymerases β, κ, and l. DNA polymerase β is capable of bypassing a psoralen cross-link with a low efficiency. Cell extracts prepared from DNA polymerase β knockout mouse embryonic fibroblasts showed a reduced bypass activity of the psoralen cross-link, and purified DNA polymerase β restored the bypass activity. In addition, DNA polymerase l misincorporated thymine across the psoralen cross-link and DNA polymerase κ extended these mispaired primer ends, suggesting that DNA polymerase l may serve as an inserter and DNA polymerase κ may play a role as an extender in the repair of psoralen DNA interstrand cross-links. The results demonstrated here indicate that multiple DNA polymerases could participate in TLS steps in mammalian DNA interstrand cross-link repair.

AB - Repair of DNA interstrand cross-links in mammalian cells involves several biochemically distinctive processes, including the release of one of the cross-linked strands and translesion DNA synthesis (TLS). In this report, we investigated the in vitro TLS activity of a psoralen DNA interstrand cross-link by three DNA repair polymerases, DNA polymerases β, κ, and l. DNA polymerase β is capable of bypassing a psoralen cross-link with a low efficiency. Cell extracts prepared from DNA polymerase β knockout mouse embryonic fibroblasts showed a reduced bypass activity of the psoralen cross-link, and purified DNA polymerase β restored the bypass activity. In addition, DNA polymerase l misincorporated thymine across the psoralen cross-link and DNA polymerase κ extended these mispaired primer ends, suggesting that DNA polymerase l may serve as an inserter and DNA polymerase κ may play a role as an extender in the repair of psoralen DNA interstrand cross-links. The results demonstrated here indicate that multiple DNA polymerases could participate in TLS steps in mammalian DNA interstrand cross-link repair.

UR - http://www.scopus.com/inward/record.url?scp=84868568928&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868568928&partnerID=8YFLogxK

U2 - 10.1021/bi3008565

DO - 10.1021/bi3008565

M3 - Article

VL - 51

SP - 8931

EP - 8938

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 44

ER -