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Silencing mediated by the Schizosaccharomyces pombe HIRA complex is dependent upon the Hpc2-like protein, Hip4
- Source :
- PLoS ONE, Vol 5, Iss 10, p e13488 (2010), Anderson, H E, Kagansky, A, Wardle, J, Rappsilber, J, Allshire, R C & Whitehall, S K 2010, ' Silencing Mediated by the Schizosaccharomyces pombe HIRA Complex Is Dependent upon the Hpc2-Like Protein, Hip4 ', PLoS ONE, vol. 5, no. 10, e13488, pp.-. https://doi.org/10.1371/journal.pone.0013488, PLoS ONE
- Publication Year :
- 2010
- Publisher :
- Public Library of Science (PLoS), 2010.
-
Abstract
- Background: HIRA (or Hir) proteins are conserved histone chaperones that function in multi-subunit complexes to mediate replication-independent nucleosome assembly. We have previously demonstrated that the Schizosaccharomyces pombe HIRA proteins, Hip1 and Slm9, form a complex with a TPR repeat protein called Hip3. Here we have identified a new subunit of this complex.Methodology/Principal Findings: To identify proteins that interact with the HIRA complex, rapid affinity purifications of Slm9 were performed. Multiple components of the chaperonin containing TCP-1 complex (CCT) and the 19S subunit of the proteasome reproducibly co-purified with Slm9, suggesting that HIRA interacts with these complexes. Slm9 was also found to interact with a previously uncharacterised protein (SPBC947.08c), that we called Hip4. Hip4 contains a HRD domain which is a characteristic of the budding yeast and human HIRA/Hir-binding proteins, Hpc2 and UBN1. Co-precipitation experiments revealed that Hip4 is stably associated with all of the other components of the HIRA complex and deletion of hip4(+) resulted in the characteristic phenotypes of cells lacking HIRA function, such as temperature sensitivity, an elongated cell morphology and hypersensitivity to the spindle poison, thiabendazole. Moreover, loss of Hip4 function alleviated the heterochromatic silencing of reporter genes located in the mating type locus and centromeres and was associated with increased levels of non-coding transcripts derived from centromeric repeat sequences. Hip4 was also found to be required for the distinct form of silencing that controls the expression of Tf2 LTR retrotransposons.Conclusions/Significance: Overall, these results indicate that Hip4 is an integral component of the HIRA complex that is required for transcriptional silencing at multiple loci.
- Subjects :
- Nucleosome assembly
Protein subunit
Molecular Sequence Data
lcsh:Medicine
Molecular Biology/Histone Modification
Cell morphology
DNA-binding protein
Chaperonin
Open Reading Frames
03 medical and health sciences
Tandem Mass Spectrometry
Genetics and Genomics/Epigenetics
Schizosaccharomyces
Amino Acid Sequence
Gene Silencing
Molecular Biology/Chromatin Structure
lcsh:Science
Cell Biology/Gene Expression
DNA Primers
030304 developmental biology
Genetics
0303 health sciences
Multidisciplinary
Base Sequence
Sequence Homology, Amino Acid
biology
Reverse Transcriptase Polymerase Chain Reaction
030302 biochemistry & molecular biology
lcsh:R
Molecular Biology/Chromosome Structure
Genetics and Genomics/Gene Expression
biology.organism_classification
Molecular Biology/Centromeres
3. Good health
Genetics and Genomics/Chromosome Biology
Histone
Schizosaccharomyces pombe
biology.protein
lcsh:Q
Schizosaccharomyces pombe Proteins
Chromatography, Liquid
Transcription Factors
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 5
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....3a95a5346e441e3720d8b5d515733c0f