Rad53 phosphorylation site clusters are important for Rad53 regulation and signaling.

Abstract:

:Budding yeast Rad53 is an essential protein kinase that is phosphorylated and activated in a MEC1- and TEL1-dependent manner in response to DNA damage. We studied the role of Rad53 phosphorylation through mutation of consensus phosphorylation sites for upstream kinases Mec1 and Tel1. Alanine substitution of the Rad53 amino-terminal TQ cluster region reduced viability and impaired checkpoint functions. These substitution mutations spared the basal interaction with Asf1 and the DNA damage-induced interactions with Rad9. However, they caused a decrease in DNA damage-induced Rad53 kinase activity and an impaired interaction with the protein kinase Dun1. The Dun1 FHA (Forkhead-associated) domain recognized the amino-terminal TQ cluster of Rad53 after DNA damage or replication blockade. Thus, the phosphorylation of Rad53 by upstream kinases is important not only for Rad53 activation but also for creation of an interface between Rad53 and Dun1.

journal_name

Mol Cell Biol

authors

Lee SJ,Schwartz MF,Duong JK,Stern DF

doi

10.1128/mcb.23.17.6300-6314.2003

subject

Has Abstract

pub_date

2003-09-01 00:00:00

pages

6300-14

issue

17

eissn

0270-7306

issn

1098-5549

journal_volume

23

pub_type

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