|
| Status |
Public on Aug 10, 2022 |
| Title |
Mutational analysis of Rap1 reveals specific vulnerability for long telomeres fusions |
| Organisms |
Escherichia coli; Schizosaccharomyces pombe |
| Experiment type |
Other
|
| Summary |
The conserved Rap1 protein is part of the shelterin complex that plays critical roles in chromosome end protection and telomere length homeostasis. Previous studies addressed how fission yeast Rap1 contributes to telomere length maintenance, but the mechanism by which the protein inhibits end fusions has remained elusive. Here, we use a genetic screen in combination with high throughput sequencing to identify several amino acid positions in Rap1 that have a key role in end protection. Interestingly, mutations at these sites render cells susceptible to genome instability in a conditional manner with longer telomeres being prone to undergoing end fusions, while short telomeres are sufficiently protected. The protection of long telomeres requires their nuclear envelope attachment mediated by the Rap1-Bqt4 interaction. Our data demonstrates that longer telomeres pose an additional challenge for the maintenance of genome integrity and provides an explanation for a species-specific upper limit in telomere length.
|
| |
|
| Overall design |
One sample of the mutanesis library from E.coli and prior to transformation to S.pombe; One sample from S.pombe after recovery from transformation; and 3 biological replicates of the time course for 5 rounds were included. The Rap1 fragment from each sample was amplified and amplicon libraries were generated for illumina sequencing. Changes of non-native nucleotides abundances at each nucelotide position were analyzed to select for amino acid positions that are important for end protection.
|
| |
|
| Contributor(s) |
Pan L, Tormey D, Bobon N, Baumann P |
| Citation(s) |
36059259 |
| |
| Submission date |
Dec 13, 2021 |
| Last update date |
Nov 02, 2022 |
| Contact name |
Peter Baumann |
| Organization name |
Johannes Gutenberg University of Mainz
|
| Street address |
Ackermannweg 4
|
| City |
Mainz |
| ZIP/Postal code |
55124 |
| Country |
Germany |
| |
|
| Platforms (2) |
| GPL17225 |
Illumina HiSeq 2500 (Schizosaccharomyces pombe) |
| GPL18133 |
Illumina HiSeq 2500 (Escherichia coli) |
|
| Samples (17)
|
|
| Relations |
| BioProject |
PRJNA788437 |
| SRA |
SRP350521 |
Less...
More...