The post-translational modifications of proliferating cell nuclear antigen: acetylation, not phosphorylation, plays an important role in the regulation of its function.
The diverse function of proliferating cell nuclear antigen (PCNA) is thought to be due, in large part, to post-translational modifications. Here we show by high resolution two-dimensional PAGE analysis that there are three distinct PCNA isoforms that differ in their acetylation status. The moderately acetylated main (M) form was found in all of the subcellular compartments of cycling cells, whereas the highly acetylated acidic form was primarily found in the nucleoplasm, nuclear matrix, and chromatin. Interestingly, the deacetylated basic form was most pronounced in the nucleoplasm of cycling cells. The cells in G(0) and the cytoplasm of cycling cells contained primarily the M form only. Because p300 and histone deacetylase (HDAC1) were co-immunoprecipitated with PCNA, they are likely responsible for the acetylation and deacetylation of PCNA, respectively. We also found that deacetylation reduced the ability of PCNA to bind to DNA polymerases beta and delta. Taken together, our data support a model where the acidic and M forms participate in DNA replication, whereas the basic form is associated with the termination of DNA replication.
Department of Research, Northeastern Ontario Regional Cancer Centre, 41 Ramsey Lake Road, Sudbury, Ontario P3E 5J1, Canada.
Stanislav N
SN
Naryzhny
Hoyun
H
Lee
D000107:Acetylation; D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000737:chemistry; D006224:Cricetinae; D003593:Cytoplasm / Q000378:metabolism; D004258:DNA Polymerase III / Q000378:metabolism; D019951:DNA Polymerase beta / Q000737:chemistry / Q000378:metabolism; D004261:DNA Replication; D015180:Electrophoresis, Gel, Two-Dimensional; D015151:Immunoblotting; D010766:Phosphorylation; D011233:Precipitin Tests; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry* / Q000235:genetics; D020033:Protein Isoforms; D011499:Protein Processing, Post-Translational; D016192:Resting Phase, Cell Cycle; D016196:S Phase; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D002843:Chromatin; D018809:Proliferating Cell Nuclear Antigen; D020033:Protein Isoforms; D004258:DNA Polymerase III; D019951:DNA Polymerase beta