Studies on Substrate Recognition by the Budding Yeast Separase
| Autor: | Nadine C.D. Hornig, Thomas Porstmann, Frank Uhlmann, Matt Sullivan |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Cohesin complex Chromosomal Proteins Non-Histone Amino Acid Motifs Molecular Sequence Data Mitosis Cell Cycle Proteins Biology Cleavage (embryo) Biochemistry Substrate Specificity Endopeptidases Consensus sequence Sister chromatids Amino Acid Sequence Kinetochores Molecular Biology Separase Anaphase Genetics Binding Sites Sequence Homology Amino Acid Kinetochore Nuclear Proteins Cell Biology Phosphoproteins Protein Structure Tertiary Cell biology Establishment of sister chromatid cohesion Mutation Saccharomycetales Mutagenesis Site-Directed Microtubule-Associated Proteins Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 279:1191-1196 |
| ISSN: | 0021-9258 |
| Popis: | Sister chromatid cohesion is resolved at anaphase onset when separase, a site-specific protease, cleaves the Scc1 subunit of the chromosomal cohesin complex that is responsible for holding sister chromatids together. This mechanism to initiate anaphase is conserved in eukaryotes from budding yeast to man. Budding yeast separase recognizes and cleaves two conserved peptide motifs within Scc1. In addition, separase cleaves a similar motif in the kinetochore and spindle protein Slk19. Separase may cleave further substrate proteins to orchestrate multiple cellular events that take place during anaphase. To investigate substrate recognition by budding yeast separase we analyzed the sequence requirements at one of the Scc1 cleavage site motifs by systematic mutagenesis. We derived a cleavage site consensus motif (not(FKRWY))(ACFHILMPVWY)(DE)X(AGSV)R/X. This motif is found in 1,139 of 5,889 predicted yeast proteins. We analyzed 28 candidate proteins containing this motif as well as 35 proteins that contain a core (DE)XXR motif. We could so far not confirm new separase substrates, but we have uncovered other forms of mitotic regulation of some of the proteins. We studied whether determinants other than the cleavage site motif mediate separase-substrate interaction. When the separase active site was occupied with a peptide inhibitor covering the cleavage site motif, separase still efficiently interacted with its substrate Scc1. This suggests that separase recognizes both a cleavage site consensus sequence as well as features outside the cleavage site. |
| Databáze: | OpenAIRE |
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