FHA domain boundaries of the dun1p and rad53p cell cycle checkpoint kinases.
Dun1p and Rad53p of the budding yeast Saccharomyces cerevisiae are members of a conserved family of cell cycle checkpoint protein kinases that contain forkhead-associated (FHA) domains. Here, we demonstrate that these FHA domains contain 130-140 residues, and are thus considerably larger than previously predicted by sequence comparisons (55-75 residues). In vivo, expression of the proteolytically defined Dun1p FHA domain, but not a fragment containing only the predicted domain boundaries, inhibited the transcriptional induction of repair genes following replication blocks. This indicates that the non-catalytic FHA domain plays an important role in the transcriptional function of the Dun1p protein kinase.
Hammet
A
A
St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, Vic., Australia.
Pike
B L
BL
Mitchelhill
K I
KI
Teh
T
T
Kobe
B
B
House
C M
CM
Kemp
B E
BE
Heierhorst
J
J
D000595:Amino Acid Sequence; D020134:Catalytic Domain / Q000235:genetics / Q000502:physiology; D018797:Cell Cycle Proteins; D064447:Checkpoint Kinase 2; D002918:Chymotrypsin / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics; D006918:Hydroxyurea / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D010446:Peptide Fragments; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D011494:Protein Kinases; C104169:DUN1 protein, S cerevisiae; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase; D006918:Hydroxyurea