Mutational analyses of the interacting domains of Schizosaccharomyces pombe Byr2 with 14-3-3s.
S. pombe
14-3-3
Byr2
MAP kinase
Rad24
Rad25
Journal
Current genetics
ISSN: 1432-0983
Titre abrégé: Curr Genet
Pays: United States
ID NLM: 8004904
Informations de publication
Date de publication:
24 Jun 2024
24 Jun 2024
Historique:
received:
04
04
2024
accepted:
16
06
2024
revised:
10
06
2024
medline:
24
6
2024
pubmed:
24
6
2024
entrez:
24
6
2024
Statut:
epublish
Résumé
The Byr2 kinase of fission yeast Schizosaccharomyces pombe is recruited to the membrane with the assistance of Ras1. Byr2 is also negatively regulated by 14-3-3 proteins encoded by rad24 and rad25. We conducted domain and mutational analysis of Byr2 to determine which region is critical for its binding to 14-3-3 proteins. Rad24 and Rad25 bound to both the Ras interaction domain in the N-terminus and to the C-terminal catalytic domain of Byr2. When amino acid residues S87 and T94 of the Ras-interacting domain of Byr2 were mutated to alanine, Rad24 could no longer bind to Byr2. S402, S566, S650, and S654 mutations in the C-terminal domain of Byr2 also abolished its interaction with Rad24 and Rad25. More than three mutations in the C-terminal domain were required to abolish completely its interaction with 14-3-3 protein, suggesting that multiple residues are involved in this interaction. Expression of the N-terminal domain of Byr2 in wild-type cells lowered the mating ratio, because it likely blocked the interaction of Byr2 with Ste4 and Ras1, whereas expression of the catalytic domain of Byr2 increased the mating ratio as a result of freeing from intramolecular regulation by the N-terminal domain of Byr2. The S87A and T94A mutations of Byr2 increased the mating ratio and attenuated inhibition of Byr2 by Rad24; therefore, these two amino acids are critical for its regulation by Rad24. S566 of Byr2 is critical for activity of Byr2 but not for its interaction with 14-3-3 proteins. In this study, we show that 14-3-3 proteins interact with two separate domains in Byr2 as negative regulators.
Identifiants
pubmed: 38913087
doi: 10.1007/s00294-024-01293-7
pii: 10.1007/s00294-024-01293-7
doi:
Substances chimiques
14-3-3 Proteins
0
Schizosaccharomyces pombe Proteins
0
rad24 protein, S pombe
0
Cell Cycle Proteins
0
Intracellular Signaling Peptides and Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
8Subventions
Organisme : Grants-in-Aid for Scientific Research , Japan
ID : #10660087 and #12660301
Informations de copyright
© 2024. The Author(s).
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