Use this URL to cite or link to this record in EThOS:
Title: Mts2+ gene of fission yeast : interactions and mutation analysis
Author: McGurk, Gordon Benedict
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1997
Availability of Full Text:
Access from EThOS:
Full text unavailable from EThOS. Please try the link below.
Access from Institution:
The primary objective of this work was the isolation and characterisation of gene products which interacted with Mts2p. This was achieved by the use of the yeast '2-hybrid' screen, and enabled isolation of a previously identified S. pombe gene let1+, which is homologous to a gene from the budding yeast S. cerevisiae. The product of this homologue, SUG1, also a member of the AAA family, was thought to be in transcription or transcriptional regulation. Characterisation of the phenotype resulting from a disruption of let1+ suggested that like mts2+, it encoded a subunit of the 26S proteasome. Further evidence is presented in favour of this argument. In addition to let1+, a novel S. pombe gene aps1+ was located. Aps1+ encodes the S. pombe homologue of the mouse MSS1 gene, which was originally isolated as a suppressor of a mutation in a yeast genes encoding a protein kinase. In this laboratory, however, MSS1 was isolated as a multicopy suppressor of the ts phenotype of mts2-1. The interactions between Mts2p, Let1P and Aps1p, the products of mts2+, let1+ and aps1+ respectively, were studied using the yeast 2-hybrid system. The region of interaction between Let1p and Mts2p were defined, and the results suggest that, as is the case for two other ATPase subunits of the 26S proteasome, the N-terminus of each protein is important in mediating this interaction. In a separate screen to look for genes which were involved in the enhancement of position effect variegation at the centromere, 4 cold sensitive (cs) alleles of mts2 were isolated. Mutation analysis was performed on these and on the three ts alleles of mts2, which had been isolated in the original drug resistance screen. All of the mutations lie in a 230 amino acid region which is highly conserved between all members of the AAA protein family. The phenotype of all of these mutants was studied with respect to morphology, DNA content and MBC resistance. The results indicate that the three ts alleles are more drug resistant and have a greater morphological deformation than the cs alleles.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available