PCNA and DNA polymerase δ catalytic subunit from Schizosaccharomyces pombe do not interact directly

Isabelle Tratner; Karine Piard; Muriel Grenon; Mylène Perderiset; Giuseppe Baldacci

doi:10.1006/bbrc.1997.6082

PCNA and DNA polymerase δ catalytic subunit from Schizosaccharomyces pombe do not interact directly

Isabelle Tratner, Karine Piard, Muriel Grenon, Mylène Perderiset, Giuseppe Baldacci

Unité Propre de Recherche

Research output: Contribution to a Journal (Peer & Non Peer) › Article › peer-review

20 Citations (Scopus)

Abstract

DNA polymerase δ (pol δ) is constituted of at least two subunits: the catalytic subunit of about 125 kDa (p125), and a subunit of approximately 50 kDa (p50) of unknown function. Processivity of pol δ is dependent on its auxiliary protein PCNA (proliferating cell nuclear antigen). Contradictory data were reported regarding a direct interaction between p125 and PCNA. We investigated this matter further using the baculovirus system to overexpress p125 and PCNA from S. pombe. We show that the recombinant p125 is active for basal DNA polymerase activity and for 3'→5' exonuclease activity but is not stimulated by PCNA. Interaction between p125 and PCNA was tested by: (i) co-immunoprecipitation assay using antibodies specific for one or other polypeptides after co-expression in insect cells, and (ii) a two-hybrid assay. In both cases, no direct interaction between the two proteins was detected. Taken together, our data show that p125 and PCNA do not interact directly.

Original language	English
Pages (from-to)	321-328
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	231
Issue number	2
DOIs	https://doi.org/10.1006/bbrc.1997.6082
Publication status	Published - 13 Feb 1997
Externally published	Yes

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title = "PCNA and DNA polymerase δ catalytic subunit from Schizosaccharomyces pombe do not interact directly",

abstract = "DNA polymerase δ (pol δ) is constituted of at least two subunits: the catalytic subunit of about 125 kDa (p125), and a subunit of approximately 50 kDa (p50) of unknown function. Processivity of pol δ is dependent on its auxiliary protein PCNA (proliferating cell nuclear antigen). Contradictory data were reported regarding a direct interaction between p125 and PCNA. We investigated this matter further using the baculovirus system to overexpress p125 and PCNA from S. pombe. We show that the recombinant p125 is active for basal DNA polymerase activity and for 3'→5' exonuclease activity but is not stimulated by PCNA. Interaction between p125 and PCNA was tested by: (i) co-immunoprecipitation assay using antibodies specific for one or other polypeptides after co-expression in insect cells, and (ii) a two-hybrid assay. In both cases, no direct interaction between the two proteins was detected. Taken together, our data show that p125 and PCNA do not interact directly.",

author = "Isabelle Tratner and Karine Piard and Muriel Grenon and Myl{\`e}ne Perderiset and Giuseppe Baldacci",

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TY - JOUR

T1 - PCNA and DNA polymerase δ catalytic subunit from Schizosaccharomyces pombe do not interact directly

AU - Tratner, Isabelle

AU - Piard, Karine

AU - Grenon, Muriel

AU - Perderiset, Mylène

AU - Baldacci, Giuseppe

PY - 1997/2/13

Y1 - 1997/2/13

N2 - DNA polymerase δ (pol δ) is constituted of at least two subunits: the catalytic subunit of about 125 kDa (p125), and a subunit of approximately 50 kDa (p50) of unknown function. Processivity of pol δ is dependent on its auxiliary protein PCNA (proliferating cell nuclear antigen). Contradictory data were reported regarding a direct interaction between p125 and PCNA. We investigated this matter further using the baculovirus system to overexpress p125 and PCNA from S. pombe. We show that the recombinant p125 is active for basal DNA polymerase activity and for 3'→5' exonuclease activity but is not stimulated by PCNA. Interaction between p125 and PCNA was tested by: (i) co-immunoprecipitation assay using antibodies specific for one or other polypeptides after co-expression in insect cells, and (ii) a two-hybrid assay. In both cases, no direct interaction between the two proteins was detected. Taken together, our data show that p125 and PCNA do not interact directly.

AB - DNA polymerase δ (pol δ) is constituted of at least two subunits: the catalytic subunit of about 125 kDa (p125), and a subunit of approximately 50 kDa (p50) of unknown function. Processivity of pol δ is dependent on its auxiliary protein PCNA (proliferating cell nuclear antigen). Contradictory data were reported regarding a direct interaction between p125 and PCNA. We investigated this matter further using the baculovirus system to overexpress p125 and PCNA from S. pombe. We show that the recombinant p125 is active for basal DNA polymerase activity and for 3'→5' exonuclease activity but is not stimulated by PCNA. Interaction between p125 and PCNA was tested by: (i) co-immunoprecipitation assay using antibodies specific for one or other polypeptides after co-expression in insect cells, and (ii) a two-hybrid assay. In both cases, no direct interaction between the two proteins was detected. Taken together, our data show that p125 and PCNA do not interact directly.

UR - https://www.scopus.com/pages/publications/0031566245

U2 - 10.1006/bbrc.1997.6082

DO - 10.1006/bbrc.1997.6082

M3 - Article

SN - 0006-291X

VL - 231

SP - 321

EP - 328

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 2

ER -

PCNA and DNA polymerase δ catalytic subunit from Schizosaccharomyces pombe do not interact directly

Abstract

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