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Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal
The HMG (high-mobility group)-box-containing chromatin-remodelling factor SRY (sex-determining region on the Y chromosome) plays a key role in sex determination. Its role in the nucleus is critically dependent on two NLSs (nuclear localization signals) that flank its HMG domain: the C-terminally loc...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2911679/ https://www.ncbi.nlm.nih.gov/pubmed/20528776 http://dx.doi.org/10.1042/BJ20091758 |
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author | Kaur, Gurpreet Delluc-Clavieres, Aurelie Poon, Ivan K. H. Forwood, Jade K. Glover, Dominic J. Jans, David A. |
author_facet | Kaur, Gurpreet Delluc-Clavieres, Aurelie Poon, Ivan K. H. Forwood, Jade K. Glover, Dominic J. Jans, David A. |
author_sort | Kaur, Gurpreet |
collection | PubMed |
description | The HMG (high-mobility group)-box-containing chromatin-remodelling factor SRY (sex-determining region on the Y chromosome) plays a key role in sex determination. Its role in the nucleus is critically dependent on two NLSs (nuclear localization signals) that flank its HMG domain: the C-terminally located ‘β-NLS’ that mediates nuclear transport through Impβ1 (importin β1) and the N-terminally located ‘CaM-NLS’ which is known to recognize the calcium-binding protein CaM (calmodulin). In the present study, we examined a number of missense mutations in the SRY CaM-NLS from human XY sex-reversed females for the first time, showing that they result in significantly reduced nuclear localization of GFP (green fluorescent protein)–SRY fusion proteins in transfected cells compared with wild-type. The CaM antagonist CDZ (calmidazolium chloride) was found to significantly reduce wild-type SRY nuclear accumulation, indicating dependence of SRY nuclear import on CaM. Intriguingly, the CaM-NLS mutants were all resistant to CDZ's effects, implying a loss of interaction with CaM, which was confirmed by direct binding experiments. CaM-binding/resultant nuclear accumulation was the only property of SRY found to be impaired by two of the CaM-NLS mutations, implying that inhibition of CaM-dependent nuclear import is the basis of sex reversal in these cases. Importantly, the CaM-NLS is conserved in other HMG-box-domain-containing proteins such as SOX-2, -9, -10 and HMGN1, all of which were found for the first time to rely on CaM for optimal nuclear localization. CaM-dependent nuclear translocation is thus a common mechanism for this family of important transcription factors. |
format | Text |
id | pubmed-2911679 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-29116792010-08-02 Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal Kaur, Gurpreet Delluc-Clavieres, Aurelie Poon, Ivan K. H. Forwood, Jade K. Glover, Dominic J. Jans, David A. Biochem J Research Article The HMG (high-mobility group)-box-containing chromatin-remodelling factor SRY (sex-determining region on the Y chromosome) plays a key role in sex determination. Its role in the nucleus is critically dependent on two NLSs (nuclear localization signals) that flank its HMG domain: the C-terminally located ‘β-NLS’ that mediates nuclear transport through Impβ1 (importin β1) and the N-terminally located ‘CaM-NLS’ which is known to recognize the calcium-binding protein CaM (calmodulin). In the present study, we examined a number of missense mutations in the SRY CaM-NLS from human XY sex-reversed females for the first time, showing that they result in significantly reduced nuclear localization of GFP (green fluorescent protein)–SRY fusion proteins in transfected cells compared with wild-type. The CaM antagonist CDZ (calmidazolium chloride) was found to significantly reduce wild-type SRY nuclear accumulation, indicating dependence of SRY nuclear import on CaM. Intriguingly, the CaM-NLS mutants were all resistant to CDZ's effects, implying a loss of interaction with CaM, which was confirmed by direct binding experiments. CaM-binding/resultant nuclear accumulation was the only property of SRY found to be impaired by two of the CaM-NLS mutations, implying that inhibition of CaM-dependent nuclear import is the basis of sex reversal in these cases. Importantly, the CaM-NLS is conserved in other HMG-box-domain-containing proteins such as SOX-2, -9, -10 and HMGN1, all of which were found for the first time to rely on CaM for optimal nuclear localization. CaM-dependent nuclear translocation is thus a common mechanism for this family of important transcription factors. Portland Press Ltd. 2010-07-28 2010-08-15 /pmc/articles/PMC2911679/ /pubmed/20528776 http://dx.doi.org/10.1042/BJ20091758 Text en © 2010 The Author(s) The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by-nc/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Kaur, Gurpreet Delluc-Clavieres, Aurelie Poon, Ivan K. H. Forwood, Jade K. Glover, Dominic J. Jans, David A. Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title | Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title_full | Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title_fullStr | Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title_full_unstemmed | Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title_short | Calmodulin-dependent nuclear import of HMG-box family nuclear factors: importance of the role of SRY in sex reversal |
title_sort | calmodulin-dependent nuclear import of hmg-box family nuclear factors: importance of the role of sry in sex reversal |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2911679/ https://www.ncbi.nlm.nih.gov/pubmed/20528776 http://dx.doi.org/10.1042/BJ20091758 |
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