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Article: Biochemical consequences of sedlin mutations that cause spondyloepiphyseal dysplasia tarda

TitleBiochemical consequences of sedlin mutations that cause spondyloepiphyseal dysplasia tarda
Authors
KeywordsBet3
Chondrocyte
Misfolding
Sedlin
Spondyloepiphyseal dysplasia tarda (SEDT)
Subcellular localization
Transport protein particle (TRAPP)
Issue Date2009
PublisherPortland Press Ltd. The Journal's web site is located at http://www.biochemj.org
Citation
Biochemical Journal, 2009, v. 423 n. 2, p. 233-242 How to Cite?
AbstractSEDT (spondyloepiphyseal dysplasia tarda) is a late-onset X-linked recessive skeletal dysplasia caused by mutations in the gene SEDL coding for sedlin. In the present paper,we investigated four missense mutations observed in SEDT and compare biochemical and cellular characteristics relative to the wild-type protein to address the mechanism of disease and to gain insight into the function of the sedlin protein. In situ hybridization and immunohistochemical experiments in mouse growth plates revealed sedlin to be predominantly expressed in proliferating and hypertrophic chondrocytes. Cell culture studies showed that the wildtype protein localized predominantly in the vicinity of the nucleus and the Golgi, with further localization around the cytoplasm, whereas mutation resulted in mislocalization. The D47Y mutant was expressed similarly to the wild-type, but the S73L, F83S and V130D mutants showed particularly low levels of expression that were rescued in the presence of the proteasome inhibitor MG132 (benzyloxycarbonyl-leucylleucylleucinal). Furthermore, whereas the D47Y mutant folded similarly and had similar stability to the wild-type sedlin as shown by CD and fluorescence, the S73L, F83S and V130D mutants all misfolded during expression. Two independent assays showed that the D47Y mutation resulted in an increased affinity for the transport protein particle component Bet3 compared with the wild-type sedlin. Our results suggest that the sedlin mutations S73L, F83S and V130D cause SEDT by sedlin misfolding, whereas the D47Y mutation may influence normal TRAPP (transport protein particle) dynamics. © The Authors Journal compilation. © 2009 Biochemical Society.
Persistent Identifierhttp://hdl.handle.net/10722/68139
ISSN
2015 Impact Factor: 3.562
2015 SCImago Journal Rankings: 2.582
ISI Accession Number ID
Funding AgencyGrant Number
Hong Kong University Grants Council
Funding Information:

This work was supported by the Hong Kong University Grants Council under the Area of Excellence Grant "Developmental Genomics and Skeletal Research".

References

 

DC FieldValueLanguage
dc.contributor.authorChoi, MYen_HK
dc.contributor.authorChan, CCYen_HK
dc.contributor.authorChan, Den_HK
dc.contributor.authorLuk, KDen_HK
dc.contributor.authorCheah, KSEen_HK
dc.contributor.authorTanner, JAen_HK
dc.date.accessioned2010-09-06T06:01:44Z-
dc.date.available2010-09-06T06:01:44Z-
dc.date.issued2009en_HK
dc.identifier.citationBiochemical Journal, 2009, v. 423 n. 2, p. 233-242en_HK
dc.identifier.issn0264-6021en_HK
dc.identifier.urihttp://hdl.handle.net/10722/68139-
dc.description.abstractSEDT (spondyloepiphyseal dysplasia tarda) is a late-onset X-linked recessive skeletal dysplasia caused by mutations in the gene SEDL coding for sedlin. In the present paper,we investigated four missense mutations observed in SEDT and compare biochemical and cellular characteristics relative to the wild-type protein to address the mechanism of disease and to gain insight into the function of the sedlin protein. In situ hybridization and immunohistochemical experiments in mouse growth plates revealed sedlin to be predominantly expressed in proliferating and hypertrophic chondrocytes. Cell culture studies showed that the wildtype protein localized predominantly in the vicinity of the nucleus and the Golgi, with further localization around the cytoplasm, whereas mutation resulted in mislocalization. The D47Y mutant was expressed similarly to the wild-type, but the S73L, F83S and V130D mutants showed particularly low levels of expression that were rescued in the presence of the proteasome inhibitor MG132 (benzyloxycarbonyl-leucylleucylleucinal). Furthermore, whereas the D47Y mutant folded similarly and had similar stability to the wild-type sedlin as shown by CD and fluorescence, the S73L, F83S and V130D mutants all misfolded during expression. Two independent assays showed that the D47Y mutation resulted in an increased affinity for the transport protein particle component Bet3 compared with the wild-type sedlin. Our results suggest that the sedlin mutations S73L, F83S and V130D cause SEDT by sedlin misfolding, whereas the D47Y mutation may influence normal TRAPP (transport protein particle) dynamics. © The Authors Journal compilation. © 2009 Biochemical Society.en_HK
dc.languageengen_HK
dc.publisherPortland Press Ltd. The Journal's web site is located at http://www.biochemj.orgen_HK
dc.relation.ispartofBiochemical Journalen_HK
dc.rightsThe final version of record is available at [Journal URL].-
dc.subjectBet3en_HK
dc.subjectChondrocyteen_HK
dc.subjectMisfoldingen_HK
dc.subjectSedlinen_HK
dc.subjectSpondyloepiphyseal dysplasia tarda (SEDT)en_HK
dc.subjectSubcellular localizationen_HK
dc.subjectTransport protein particle (TRAPP)en_HK
dc.subject.meshAmino Acid Substitution - genetics - physiology-
dc.subject.meshMembrane Transport Proteins - chemistry - genetics - metabolism - physiology-
dc.subject.meshMutation - physiology-
dc.subject.meshOsteochondrodysplasias - genetics-
dc.subject.meshTranscription Factors - chemistry - genetics - metabolism - physiology-
dc.titleBiochemical consequences of sedlin mutations that cause spondyloepiphyseal dysplasia tardaen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0264-6021&volume=423&issue=2&spage=233&epage=242&date=2009&atitle=Biochemical+consequences+of+sedlin+mutations+that+cause+spondyloepiphyseal+dysplasia+tardaen_HK
dc.identifier.emailChan, D:chand@hkucc.hku.hken_HK
dc.identifier.emailLuk, KD:hcm21000@hku.hken_HK
dc.identifier.emailCheah, KSE:hrmbdkc@hku.hken_HK
dc.identifier.emailTanner, JA:jatanner@hku.hken_HK
dc.identifier.authorityChan, D=rp00540en_HK
dc.identifier.authorityLuk, KD=rp00333en_HK
dc.identifier.authorityCheah, KSE=rp00342en_HK
dc.identifier.authorityTanner, JA=rp00495en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1042/BJ20090541en_HK
dc.identifier.pmid19650763-
dc.identifier.scopuseid_2-s2.0-70350109531en_HK
dc.identifier.hkuros163946en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-70350109531&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume423en_HK
dc.identifier.issue2en_HK
dc.identifier.spage233en_HK
dc.identifier.epage242en_HK
dc.identifier.eissn1470-8728-
dc.identifier.isiWOS:000271067700009-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridChoi, MY=8309672900en_HK
dc.identifier.scopusauthoridChan, CCY=36984607300en_HK
dc.identifier.scopusauthoridChan, D=7402216545en_HK
dc.identifier.scopusauthoridLuk, KD=7201921573en_HK
dc.identifier.scopusauthoridCheah, KSE=35387746200en_HK
dc.identifier.scopusauthoridTanner, JA=35513993000en_HK

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