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- Publisher Website: 10.1016/j.bbrc.2010.09.049
- Scopus: eid_2-s2.0-77957762067
- PMID: 20850414
- WOS: WOS:000283571700019
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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
Title | 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 | ||||||
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Authors | |||||||
Keywords | Acrosome Immunoprecipitation Perinuclei Spermatogenesis | ||||||
Issue Date | 2010 | ||||||
Publisher | Academic 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? | ||||||
Abstract | VAD1.3 (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 Identifier | http://hdl.handle.net/10722/139906 | ||||||
ISSN | 2023 Impact Factor: 2.5 2023 SCImago Journal Rankings: 0.770 | ||||||
ISI Accession Number ID |
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 Field | Value | Language |
---|---|---|
dc.contributor.author | Zuo, Y | en_HK |
dc.contributor.author | Gao, J | en_HK |
dc.contributor.author | Yeung, WSB | en_HK |
dc.contributor.author | Lee, KF | en_HK |
dc.date.accessioned | 2011-09-23T06:00:02Z | - |
dc.date.available | 2011-09-23T06:00:02Z | - |
dc.date.issued | 2010 | en_HK |
dc.identifier.citation | Biochemical And Biophysical Research Communications, 2010, v. 401 n. 2, p. 275-280 | en_HK |
dc.identifier.issn | 0006-291X | en_HK |
dc.identifier.uri | http://hdl.handle.net/10722/139906 | - |
dc.description.abstract | VAD1.3 (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.language | eng | en_US |
dc.publisher | Academic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description | en_HK |
dc.relation.ispartof | Biochemical and Biophysical Research Communications | en_HK |
dc.subject | Acrosome | en_HK |
dc.subject | Immunoprecipitation | en_HK |
dc.subject | Perinuclei | en_HK |
dc.subject | Spermatogenesis | en_HK |
dc.subject.mesh | Actins - metabolism | - |
dc.subject.mesh | Cell Nucleus - metabolism | - |
dc.subject.mesh | Golgi Apparatus - metabolism | - |
dc.subject.mesh | Proteins - genetics - metabolism | - |
dc.subject.mesh | Syntaxin 1 - metabolism | - |
dc.title | 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 | en_HK |
dc.type | Article | en_HK |
dc.identifier.email | Yeung, WSB:wsbyeung@hkucc.hku.hk | en_HK |
dc.identifier.email | Lee, KF:ckflee@hku.hk | en_HK |
dc.identifier.authority | Yeung, WSB=rp00331 | en_HK |
dc.identifier.authority | Lee, KF=rp00458 | en_HK |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.bbrc.2010.09.049 | en_HK |
dc.identifier.pmid | 20850414 | - |
dc.identifier.scopus | eid_2-s2.0-77957762067 | en_HK |
dc.identifier.hkuros | 196026 | en_US |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-77957762067&selection=ref&src=s&origin=recordpage | en_HK |
dc.identifier.volume | 401 | en_HK |
dc.identifier.issue | 2 | en_HK |
dc.identifier.spage | 275 | en_HK |
dc.identifier.epage | 280 | en_HK |
dc.identifier.isi | WOS:000283571700019 | - |
dc.publisher.place | United States | en_HK |
dc.identifier.scopusauthorid | Zuo, Y=37022027700 | en_HK |
dc.identifier.scopusauthorid | Gao, J=37021453600 | en_HK |
dc.identifier.scopusauthorid | Yeung, WSB=7102370745 | en_HK |
dc.identifier.scopusauthorid | Lee, KF=26643097500 | en_HK |
dc.identifier.citeulike | 7892953 | - |
dc.identifier.issnl | 0006-291X | - |