title: The G2/M DNA damage checkpoint inhibits mitosis through Tyr15 phosphorylation of p34cdc2 in Aspergillus nidulans.
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.
authors.affiliation: The Weis Center for Research, Geisinger Clinic, Danville, PA 17822, USA.
authors.forename: X S
authors.initials: XS
authors.lastname: Ye
authors.forename: R R
authors.initials: RR
authors.lastname: Fincher
authors.forename: A
authors.initials: A
authors.lastname: Tang
authors.forename: S A
authors.initials: SA
authors.lastname: Osmani
mesh_terms: D001233:Aspergillus nidulans / Q000166:cytology* / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018797:Cell Cycle Proteins; D004249:DNA Damage; D004260:DNA Repair; D004271:DNA, Fungal / Q000187:drug effects / Q000235:genetics; D016195:G2 Phase / Q000235:genetics; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; D010766:Phosphorylation / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D009153:Mutagens; D014443:Tyrosine; D008741:Methyl Methanesulfonate; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; C442331:NIMA-related kinase 6; D017346:Protein Serine-Threonine Kinases; D016203:CDC2 Protein Kinase