Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair.
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 (...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3699651?pdf=render |
| _version_ | 1811228035381198848 |
|---|---|
| author | Tiago J Dantas Owen M Daly Pauline C Conroy Martin Tomas Yifan Wang Pierce Lalor Peter Dockery Elisa Ferrando-May Ciaran G Morrison |
| author_facet | Tiago J Dantas Owen M Daly Pauline C Conroy Martin Tomas Yifan Wang Pierce Lalor Peter Dockery Elisa Ferrando-May Ciaran G Morrison |
| author_sort | Tiago J Dantas |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T09:51:43Z |
| format | Article |
| id | doaj.art-179396f4c2624220b6dd61dd005a94f3 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T09:51:43Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-179396f4c2624220b6dd61dd005a94f32022-12-22T03:37:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0187e6848710.1371/journal.pone.0068487Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair.Tiago J DantasOwen M DalyPauline C ConroyMartin TomasYifan WangPierce LalorPeter DockeryElisa Ferrando-MayCiaran G MorrisonCentrosomes, 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.http://europepmc.org/articles/PMC3699651?pdf=render |
| spellingShingle | Tiago J Dantas Owen M Daly Pauline C Conroy Martin Tomas Yifan Wang Pierce Lalor Peter Dockery Elisa Ferrando-May Ciaran G Morrison Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. PLoS ONE |
| title | Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. |
| title_full | Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. |
| title_fullStr | Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. |
| title_full_unstemmed | Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. |
| title_short | Calcium-binding capacity of centrin2 is required for linear POC5 assembly but not for nucleotide excision repair. |
| title_sort | calcium binding capacity of centrin2 is required for linear poc5 assembly but not for nucleotide excision repair |
| url | http://europepmc.org/articles/PMC3699651?pdf=render |
| work_keys_str_mv | AT tiagojdantas calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT owenmdaly calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT paulinecconroy calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT martintomas calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT yifanwang calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT piercelalor calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT peterdockery calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT elisaferrandomay calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair AT ciarangmorrison calciumbindingcapacityofcentrin2isrequiredforlinearpoc5assemblybutnotfornucleotideexcisionrepair |