PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching

Masataka Tsuda, Saki Ogawa, Masato Ooka, Kaori Kobayashi, Kouji Hirota, Mitsuo Wakasugi, Tsukasa Matsunaga, Tetsushi Sakuma, Takashi Yamamoto, Shunsuke Chikuma, Hiroyuki Sasanuma, Michelle Debatisse, Aidan J. Doherty, Robert P. Fuchs, Shunichi Takeda

Research output: Contribution to journalArticle

Abstract

Replicative DNA polymerases are frequently stalled at damaged template strands. Stalled replication forks are restored by the DNA damage tolerance (DDT) pathways, error-prone translesion DNA synthesis (TLS) to cope with excessive DNA damage, and error-free template switching (TS) by homologous DNA recombination. PDIP38 (Pol-delta interacting protein of 38 kDa), also called Pol δ-interacting protein 2 (PolDIP2), physically associates with TLS DNA polymerases, polymerase η (Polη), Polλ, and PrimPol, and activates them in vitro. It remains unclear whether PDIP38 promotes TLS in vivo, since no method allows for measuring individual TLS events in mammalian cells. We disrupted the PDIP38 gene, generating PDIP38 -/- cells from the chicken DT40 and human TK6 B cell lines. These PDIP38 -/- cells did not show a significant sensitivity to either UV or H 2 O 2 , a phenotype not seen in any TLS-polymerase-deficient DT40 or TK6 mutants. DT40 provides a unique opportunity of examining individual TLS and TS events by the nucleotide sequence analysis of the immunoglobulin variable (Ig V) gene as the cells continuously diversify Ig V by TLS (non-templated Ig V hypermutation) and TS (Ig gene conversion) during in vitro culture. PDIP38 -/- cells showed a shift in Ig V diversification from TLS to TS. We measured the relative usage of TLS and TS in TK6 cells at a chemically synthesized UV damage (CPD) integrated into genomic DNA. The loss of PDIP38 also caused an increase in the relative usage of TS. The number of UV-induced sister chromatid exchanges, TS events associated with crossover, was increased a few times in PDIP38 -/- human and chicken cells. Collectively, the loss of PDIP38 consistently causes a shift in DDT from TLS to TS without enhancing cellular sensitivity to DNA damage. We propose that PDIP38 controls the relative usage of TLS and TS increasing usage of TLS without changing the overall capability of DDT.

Original languageEnglish
Article numbere0213383
JournalPloS one
Volume14
Issue number3
DOIs
Publication statusPublished - 2019 Mar 1

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Damage tolerance
DNA damage
DNA Damage
synthesis
DNA
Proteins
proteins
immunoglobulins
DNA-Directed DNA Polymerase
Immunoglobulins
Immunoglobulin Genes
cells
DNA-directed DNA polymerase
delta protein
Chickens
pol Genes
chickens
Gene Conversion
Sister Chromatid Exchange
Homologous Recombination

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching. / Tsuda, Masataka; Ogawa, Saki; Ooka, Masato; Kobayashi, Kaori; Hirota, Kouji; Wakasugi, Mitsuo; Matsunaga, Tsukasa; Sakuma, Tetsushi; Yamamoto, Takashi; Chikuma, Shunsuke; Sasanuma, Hiroyuki; Debatisse, Michelle; Doherty, Aidan J.; Fuchs, Robert P.; Takeda, Shunichi.

In: PloS one, Vol. 14, No. 3, e0213383, 01.03.2019.

Research output: Contribution to journalArticle

Tsuda, M, Ogawa, S, Ooka, M, Kobayashi, K, Hirota, K, Wakasugi, M, Matsunaga, T, Sakuma, T, Yamamoto, T, Chikuma, S, Sasanuma, H, Debatisse, M, Doherty, AJ, Fuchs, RP & Takeda, S 2019, 'PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching', PloS one, vol. 14, no. 3, e0213383. https://doi.org/10.1371/journal.pone.0213383
Tsuda, Masataka ; Ogawa, Saki ; Ooka, Masato ; Kobayashi, Kaori ; Hirota, Kouji ; Wakasugi, Mitsuo ; Matsunaga, Tsukasa ; Sakuma, Tetsushi ; Yamamoto, Takashi ; Chikuma, Shunsuke ; Sasanuma, Hiroyuki ; Debatisse, Michelle ; Doherty, Aidan J. ; Fuchs, Robert P. ; Takeda, Shunichi. / PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching. In: PloS one. 2019 ; Vol. 14, No. 3.
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AU - Hirota, Kouji

AU - Wakasugi, Mitsuo

AU - Matsunaga, Tsukasa

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AU - Chikuma, Shunsuke

AU - Sasanuma, Hiroyuki

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