Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers: Analysis by polyacrylamide gel electrophoresis-band mobility shift assay

Lily Ng; Maeve McConnell; Cheng Keat Tan; Kathleen M. Downey; Paul A. Fisher

doi:10.1016/s0021-9258(19)38687-9

Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers: Analysis by polyacrylamide gel electrophoresis-band mobility shift assay

Lily Ng
, Maeve McConnell
, Cheng Keat Tan
, Kathleen M. Downey
, Paul A. Fisher

Pharmacological Sciences

Stony Brook University
University of Miami

Research output: Contribution to journal › Article › peer-review

36 Scopus citations

Abstract

A polyacrylamide gel electrophoresis band-mobility shift assay was developed to study the binding of synthetic oligonucleotides by DNA polymerase δ (pol δ) and proliferating cell nuclear antigen (PCNA). As measured by this assay, neither calf thymus pol δ core enzyme nor PCNA alone bind DNA stably. However, mammalian PCNA but not Drosophila PCNA promotes the formation of a distinct pol δ·PCNA·template-primer complex. Appearance of this complex is primer-dependent but does not require Mg²⁺. Complex stability is also influenced by the presence or absence of individual dNTPs. A model for the ordered sequential interaction of pol δ, PCNA, and DNA template-primers is proposed.

Original language	English
Pages (from-to)	13571-13576
Number of pages	6
Journal	Journal of Biological Chemistry
Volume	268
Issue number	18
DOIs	https://doi.org/10.1016/s0021-9258(19)38687-9
State	Published - Jun 25 1993

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10.1016/s0021-9258(19)38687-9

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title = "Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers: Analysis by polyacrylamide gel electrophoresis-band mobility shift assay",

abstract = "A polyacrylamide gel electrophoresis band-mobility shift assay was developed to study the binding of synthetic oligonucleotides by DNA polymerase δ (pol δ) and proliferating cell nuclear antigen (PCNA). As measured by this assay, neither calf thymus pol δ core enzyme nor PCNA alone bind DNA stably. However, mammalian PCNA but not Drosophila PCNA promotes the formation of a distinct pol δ·PCNA·template-primer complex. Appearance of this complex is primer-dependent but does not require Mg2+. Complex stability is also influenced by the presence or absence of individual dNTPs. A model for the ordered sequential interaction of pol δ, PCNA, and DNA template-primers is proposed.",

author = "Lily Ng and Maeve McConnell and Tan, \{Cheng Keat\} and Downey, \{Kathleen M.\} and Fisher, \{Paul A.\}",

year = "1993",

month = jun,

day = "25",

doi = "10.1016/s0021-9258(19)38687-9",

language = "English",

volume = "268",

pages = "13571--13576",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

number = "18",

}

Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers: Analysis by polyacrylamide gel electrophoresis-band mobility shift assay. / Ng, Lily; McConnell, Maeve; Tan, Cheng Keat et al.
In: Journal of Biological Chemistry, Vol. 268, No. 18, 25.06.1993, p. 13571-13576.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers

T2 - Analysis by polyacrylamide gel electrophoresis-band mobility shift assay

AU - Ng, Lily

AU - McConnell, Maeve

AU - Tan, Cheng Keat

AU - Downey, Kathleen M.

AU - Fisher, Paul A.

PY - 1993/6/25

Y1 - 1993/6/25

N2 - A polyacrylamide gel electrophoresis band-mobility shift assay was developed to study the binding of synthetic oligonucleotides by DNA polymerase δ (pol δ) and proliferating cell nuclear antigen (PCNA). As measured by this assay, neither calf thymus pol δ core enzyme nor PCNA alone bind DNA stably. However, mammalian PCNA but not Drosophila PCNA promotes the formation of a distinct pol δ·PCNA·template-primer complex. Appearance of this complex is primer-dependent but does not require Mg2+. Complex stability is also influenced by the presence or absence of individual dNTPs. A model for the ordered sequential interaction of pol δ, PCNA, and DNA template-primers is proposed.

AB - A polyacrylamide gel electrophoresis band-mobility shift assay was developed to study the binding of synthetic oligonucleotides by DNA polymerase δ (pol δ) and proliferating cell nuclear antigen (PCNA). As measured by this assay, neither calf thymus pol δ core enzyme nor PCNA alone bind DNA stably. However, mammalian PCNA but not Drosophila PCNA promotes the formation of a distinct pol δ·PCNA·template-primer complex. Appearance of this complex is primer-dependent but does not require Mg2+. Complex stability is also influenced by the presence or absence of individual dNTPs. A model for the ordered sequential interaction of pol δ, PCNA, and DNA template-primers is proposed.

UR - https://www.scopus.com/pages/publications/0027425441

U2 - 10.1016/s0021-9258(19)38687-9

DO - 10.1016/s0021-9258(19)38687-9

M3 - Article

C2 - 8099910

AN - SCOPUS:0027425441

SN - 0021-9258

VL - 268

SP - 13571

EP - 13576

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 18

ER -

Interaction of DNA polymerase δ, proliferating cell nuclear antigen, and synthetic oligonucleotide template-primers: Analysis by polyacrylamide gel electrophoresis-band mobility shift assay

Abstract

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