Functional cdc25C dual-specificity phosphatase is required for S-phase entry in human cells.

Abstract:

:In view of the common regulatory mechanism that induces transcription of the mitotic phosphatase cdc25C and cyclin A at the beginning of S-phase, we investigated whether cdc25C was required for S-phase transit. Here, we show that in both nontransformed human fibroblasts and HeLa cells, cdc25C protein levels significantly increased concomitant with S-phase onset and cyclin A synthesis. Activity measurements on immunoprecipitates from synchronized HeLa cells revealed a sharp rise in cdc25C-associated phosphatase activity that coincided with S-phase. Microinjection of various antisense-cdc25C molecules led to inhibition of DNA synthesis in both HeLa cells and human fibroblasts. Furthermore, transfection of small interfering RNA directed against cdc25C specifically depleted cdc25C in HeLa cells without affecting cdc25A or cdc25B levels. Cdc25C RNA interference was also accompanied by S-phase inhibition. In cells depleted of cdc25C by antisense or siRNA, normal cell cycle progression could be re-established through microinjection of wild-type cdc25C protein but not inactive C377S mutant protein. Taken together, these results show that cdc25C not only plays a role at the G2/M transition but also in the modulation of DNA replication where its function is distinct from that of cdc25A.

journal_name

Mol Biol Cell

authors

Turowski P,Franckhauser C,Morris MC,Vaglio P,Fernandez A,Lamb NJ

doi

10.1091/mbc.e02-08-0515

keywords:

subject

Has Abstract

pub_date

2003-07-01 00:00:00

pages

2984-98

issue

7

eissn

1059-1524

issn

1939-4586

pii

E02-08-0515

journal_volume

14

pub_type

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