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Article: The testis-specific VAD1.3/AEP1 interacts with β-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence

TitleThe testis-specific VAD1.3/AEP1 interacts with β-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence
Authors
KeywordsAcrosome
Immunoprecipitation
Perinuclei
Spermatogenesis
Issue Date2010
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
Citation
Biochemical And Biophysical Research Communications, 2010, v. 401 n. 2, p. 275-280 How to Cite?
AbstractVAD1.3 (AEP1), a novel testis-specific gene, was first isolated from the testis of a retinol-treated vitamin-A-deficient (VAD) rat model. It is expressed at the acrosomal region of spermatids from postnatal day 25. VAD1.3 immunoreactivity is present in rat, human, monkey and porcine spermatids and spermatozoa, suggesting that VAD1.3 may play a role in acrosome formation. However, direct evidence on the detailed sub-cellular localization of the VAD1.3 protein in the acrosome and how VAD1.3 is involved in acrosome formation remains largely unknown. Here, we isolated and identified VAD1.3 interacting proteins by immunoprecipitation followed by mass spectrometry, and determined the functional motifs of VAD1.3 that were important for its specific sub-cellular location in vitro. We found that VAD1.3 bound to syntaxin 1 and β-actin proteins in vitro. Immunogold electron microscopic study localized VAD1.3 immunoreactivity to the acrosome membranes and matrix, and colocalized it with the β-actin protein. The full-length GFP-VAD (1-3601) and GFP-VAD (1-730) fusion proteins that contain the bipartite nucleus localization (BNL) signal were located in the peri-nucleus/Golgi of the transfected cells. In addition, the GFP signal colocalized with the endoplasmic reticulum marker and the syntaxin 1 protein in the transfected HeLa and GC-2spd cells. The C-terminal GFP-VAD (1770-3601) was expressed in the nucleus. Taken together, VAD1.3 interacts with β-actin and syntaxin 1 in vitro. The BNL signal may mediate the peri-nuclei localization of the protein that may interact with syntaxin 1 and β-actin for acrosome formation in spermatogenesis. © 2010.
Persistent Identifierhttp://hdl.handle.net/10722/139906
ISSN
2015 Impact Factor: 2.371
2015 SCImago Journal Rankings: 1.152
ISI Accession Number ID
Funding AgencyGrant Number
Committee on Research and Conference Grants of The University of Hong Kong
Hong Kong Research Grants Council (GRF)HKU7537/05M
Funding Information:

This work was supported in part by grants from the Committee on Research and Conference Grants of The University of Hong Kong and the Hong Kong Research Grants Council (GRF grant HKU7537/05M) to KF Lee.

References

 

DC FieldValueLanguage
dc.contributor.authorZuo, Yen_HK
dc.contributor.authorGao, Jen_HK
dc.contributor.authorYeung, WSBen_HK
dc.contributor.authorLee, KFen_HK
dc.date.accessioned2011-09-23T06:00:02Z-
dc.date.available2011-09-23T06:00:02Z-
dc.date.issued2010en_HK
dc.identifier.citationBiochemical And Biophysical Research Communications, 2010, v. 401 n. 2, p. 275-280en_HK
dc.identifier.issn0006-291Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/139906-
dc.description.abstractVAD1.3 (AEP1), a novel testis-specific gene, was first isolated from the testis of a retinol-treated vitamin-A-deficient (VAD) rat model. It is expressed at the acrosomal region of spermatids from postnatal day 25. VAD1.3 immunoreactivity is present in rat, human, monkey and porcine spermatids and spermatozoa, suggesting that VAD1.3 may play a role in acrosome formation. However, direct evidence on the detailed sub-cellular localization of the VAD1.3 protein in the acrosome and how VAD1.3 is involved in acrosome formation remains largely unknown. Here, we isolated and identified VAD1.3 interacting proteins by immunoprecipitation followed by mass spectrometry, and determined the functional motifs of VAD1.3 that were important for its specific sub-cellular location in vitro. We found that VAD1.3 bound to syntaxin 1 and β-actin proteins in vitro. Immunogold electron microscopic study localized VAD1.3 immunoreactivity to the acrosome membranes and matrix, and colocalized it with the β-actin protein. The full-length GFP-VAD (1-3601) and GFP-VAD (1-730) fusion proteins that contain the bipartite nucleus localization (BNL) signal were located in the peri-nucleus/Golgi of the transfected cells. In addition, the GFP signal colocalized with the endoplasmic reticulum marker and the syntaxin 1 protein in the transfected HeLa and GC-2spd cells. The C-terminal GFP-VAD (1770-3601) was expressed in the nucleus. Taken together, VAD1.3 interacts with β-actin and syntaxin 1 in vitro. The BNL signal may mediate the peri-nuclei localization of the protein that may interact with syntaxin 1 and β-actin for acrosome formation in spermatogenesis. © 2010.en_HK
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/descriptionen_HK
dc.relation.ispartofBiochemical and Biophysical Research Communicationsen_HK
dc.subjectAcrosomeen_HK
dc.subjectImmunoprecipitationen_HK
dc.subjectPerinucleien_HK
dc.subjectSpermatogenesisen_HK
dc.subject.meshActins - metabolism-
dc.subject.meshCell Nucleus - metabolism-
dc.subject.meshGolgi Apparatus - metabolism-
dc.subject.meshProteins - genetics - metabolism-
dc.subject.meshSyntaxin 1 - metabolism-
dc.titleThe testis-specific VAD1.3/AEP1 interacts with β-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequenceen_HK
dc.typeArticleen_HK
dc.identifier.emailYeung, WSB:wsbyeung@hkucc.hku.hken_HK
dc.identifier.emailLee, KF:ckflee@hku.hken_HK
dc.identifier.authorityYeung, WSB=rp00331en_HK
dc.identifier.authorityLee, KF=rp00458en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.bbrc.2010.09.049en_HK
dc.identifier.pmid20850414-
dc.identifier.scopuseid_2-s2.0-77957762067en_HK
dc.identifier.hkuros196026en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77957762067&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume401en_HK
dc.identifier.issue2en_HK
dc.identifier.spage275en_HK
dc.identifier.epage280en_HK
dc.identifier.isiWOS:000283571700019-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridZuo, Y=37022027700en_HK
dc.identifier.scopusauthoridGao, J=37021453600en_HK
dc.identifier.scopusauthoridYeung, WSB=7102370745en_HK
dc.identifier.scopusauthoridLee, KF=26643097500en_HK
dc.identifier.citeulike7892953-

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