PloS one, 2013-07, Vol.8 (7), p.e68487-e68487
2013
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair
- Ist Teil von
- PloS one, 2013-07, Vol.8 (7), p.e68487-e68487
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2013
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Centrosomes, the principal microtubule-organising centres in animal cells, contain centrins, small, conserved calcium-binding proteins unique to eukaryotes. Centrin2 binds to xeroderma pigmentosum group C protein (XPC), stabilising it, and its presence slightly increases nucleotide excision repair (NER) activity in vitro. In previous work, we deleted all three centrin isoforms present in chicken DT40 cells and observed delayed repair of UV-induced DNA lesions, but no centrosome abnormalities. Here, we explore how centrin2 controls NER. In the centrin null cells, we expressed centrin2 mutants that cannot bind calcium or that lack sites for phosphorylation by regulatory kinases. Expression of any of these mutants restored the UV sensitivity of centrin null cells to normal as effectively as expression of wild-type centrin. However, calcium-binding-deficient and T118A mutants showed greatly compromised localisation to centrosomes. XPC recruitment to laser-induced UV-like lesions was only slightly slower in centrin-deficient cells than in controls, and levels of XPC and its partner HRAD23B were unaffected by centrin deficiency. Interestingly, we found that overexpression of the centrin interactor POC5 leads to the assembly of linear, centrin-dependent structures that recruit other centrosomal proteins such as PCM-1 and NEDD1. Together, these observations suggest that assembly of centrins into complex structures requires calcium binding capacity, but that such assembly is not required for centrin activity in NER.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0068487Titel-ID: cdi_plos_journals_1426970421
- Format
- –
- Schlagworte
- Abnormalities, Animals, Assembly, Autoantigens - genetics, Autoantigens - metabolism, Binding, Binding proteins, Biology, C protein, Calcium, Calcium - metabolism, Calcium-Binding Proteins - genetics, Calcium-Binding Proteins - metabolism, Carrier Proteins - genetics, Carrier Proteins - metabolism, Cell cycle, Cell Cycle Proteins - genetics, Cell Cycle Proteins - metabolism, Cell Line, Cell Line, Tumor, Centrosome - metabolism, Centrosome - ultrastructure, Centrosomes, Chickens, Chlamydomonas, Chromosomes, Deoxyribonucleic acid, DNA, DNA Damage, DNA Repair, DNA-Binding Proteins - genetics, DNA-Binding Proteins - metabolism, Eukaryotes, Genetic aspects, Green Fluorescent Proteins - genetics, Green Fluorescent Proteins - metabolism, Humans, Immunoblotting, Isoforms, Kinases, Lesions, Localization, Microscopy, Electron, Transmission, Microscopy, Fluorescence, Microtubule-Associated Proteins - genetics, Microtubule-Associated Proteins - metabolism, Mutagenesis, Mutants, Nucleotide excision repair, Null cells, Phosphorylation, Phosphotransferases, Physiological aspects, Point Mutation, Protein Binding, Proteins, Recruitment, Repair, U.V. radiation, Ultraviolet radiation, Xeroderma pigmentosum, XPC protein