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Association of proliferating cell nuclear antigen with cyclin-dependent
kinases and cyclins in normal and transformed human T lymphocytes
A Szepesi, EW Gelfand and JJ Lucas
Department of Pediatrics, National Jewish Center for Immunology and
Respiratory Medicine, Denver, CO 80206.
The proliferating cell nuclear antigen (PCNA) is an auxiliary protein of
DNA polymerase delta and appears to be needed for both DNA synthesis and
DNA repair. It is present in low amount in resting normal human T
lymphocytes and, upon mitogenic stimulation with phorbol dibutyrate and
ionomycin, begins to increase in mid-G1 phase, approximately 12 to 15 hours
before entry into S phase. PCNA continues to increase in amount throughout
the cell cycle and remains high in proliferating cultures. PCNA was
extracted from activated normal T cells and from the transformed
T-lymphoblastoid cell line Jurkat by a method that recovered approximately
98% of total cellular PCNA but yet retained its associations with other
proteins. PCNA immunoprecipitates possessed H1 histone kinase activity,
which increased in parallel with increasing cellular content of PCNA. Both
the cdc2 and cdk2 kinases were found associated with PCNA in normal T
cells, in amounts consistent with detected kinase activity. The results
indicate that PCNA is not an inhibitory molecule of cdk/cyclin activity.
Both normal and transformed T cells contained PCNA in association with
cdk2, cdk4, cdk5, and cdk6, with the amount of PCNA associated with these
molecules increasing in the order listed. Relatively high amounts of PCNA
were also found associated with cyclins D2 and D3, the major cyclin
partners of cdk6 in T cells. Though detected in normal cells, PCNA/cdc2
complexes were present in exceedingly low amount, if at all, in Jurkat
cells. This cell line appeared to contain more of nearly all of the cdk and
cyclin molecules analyzed, but there seemed to be little difference in the
patterns of association of these molecules with PCNA in the cell line as
compared with normal human T cells.
Volume 84,
Issue 10,
pp. 3413-3421,
11/15/1994
Copyright © 1994 by The American Society of Hematology
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