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PMID: 9009279 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The G2/M DNA damage checkpoint inhibits mitosis through Tyr15 phosphorylation of p34cdc2 in Aspergillus nidulans.

The EMBO journal ·Vol. 16 ·No. 1 ·1997-01-02 ·Pages 182-92

Ye XS, Fincher RR, Tang A, Osmani SA

Abstract

It is possible to cause G2 arrest in Aspergillus nidulans by inactivating either p34cdc2 or NIMA. We therefore investigated the negative control of these two mitosis-promoting kinases after DNA damage. DNA damage caused rapid Tyr15 phosphorylation of p34cdc2 and transient cell cycle arrest but had little effect on the activity of NIMA. Dividing cells deficient in Tyr15 phosphorylation of p34cdc2 were sensitive to both MMS and UV irradiation and entered lethal premature mitosis with damaged DNA. However, non-dividing quiescent conidiospores of the Tyr15 mutant strain were not sensitive to DNA damage. The UV and MMS sensitivity of cells unable to tyrosine phosphorylate p34cdc2 is therefore caused by defects in DNA damage checkpoint regulation over mitosis. Both the nimA5 and nimT23 temperature-sensitive mutations cause an arrest in G2 at 42 degrees C. Addition of MMS to nimT23 G2-arrested cells caused a marked delay in their entry into mitosis upon downshift to 32 degrees C and this delay was correlated with a long delay in the dephosphorylation and activation of p34cdc2. Addition of MMS to nimA5 G2-arrested cells caused inactivation of the H1 kinase activity of p34cdc2 due to an increase in its Tyr15 phosphorylation level and delayed entry into mitosis upon return to 32 degrees C. However, if Tyr15 phosphorylation of p34cdc2 was prevented then its H1 kinase activity was not inactivated upon MMS addition to nimA5 G2-arrested cells and they rapidly progressed into a lethal mitosis upon release to 32 degrees C. Thus, Tyr15 phosphorylation of p34cdc2 in G2 arrests initiation of mitosis after DNA damage in A. nidulans.

MeSH Terms
Aspergillus nidulans/cytology,drug effects,genetics,radiation effects CDC2 Protein Kinase/antagonists & inhibitors,metabolism Cell Cycle Proteins DNA Damage DNA Repair DNA, Fungal/drug effects,genetics G2 Phase/genetics Methyl Methanesulfonate/pharmacology Mitosis/drug effects,genetics,radiation effects Mutagens/pharmacology Mutation NIMA-Related Kinase 1 NIMA-Related Kinases Phosphorylation/drug effects Protein Serine-Threonine Kinases/genetics,metabolism Tyrosine/metabolism Ultraviolet Rays
Chemicals
Cell Cycle Proteins DNA, Fungal Mutagens Tyrosine Methyl Methanesulfonate NIMA-Related Kinase 1 NIMA-Related Kinases NIMA-related kinase 6 Protein Serine-Threonine Kinases CDC2 Protein Kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ye X S
The Weis Center for Research, Geisinger Clinic, Danville, PA 17822, USA.
Fincher R R
Tang A
Osmani S A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.1.182
Published
1997-01-02
Pages
182-92
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1169625
Subset
IM
Grants
NIGMS NIH HHS · GM42564 · United States
External Links
PubMed source DOI: 10.1093/emboj/16.1.182 PMC Full Text View journal details
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