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

Abstract

Research highlights: {yields} VAD1.3 interacts {beta}-actin and syntaxin 1. {yields} VAD1.3 colocalizes {beta}-actin in spermatids. {yields} The bipartite nucleus localization (BNL) signal is important for peri-nuclear/Golgi expression in transfected cells. {yields} The C-terminal region of VAD1.3 direct nuclei localization. -- 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 {beta}-actin proteins in vitro. Immunogold electron microscopic study localized VAD1.3 immunoreactivity to the acrosome membranes and matrix, and colocalized it with the {beta}-actin protein. The full-length GFP-VAD (1-3601) and GFP-VAD (1-730) fusion proteins that contain the bipartite nucleusmore » 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 {beta}-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 {beta}-actin for acrosome formation in spermatogenesis.« less

Authors:
;  [1];  [1];  [1]
  1. Department of Obstetrics and Gynaecology, The University of Hong Kong, Pokfulam (Hong Kong)
Publication Date:
OSTI Identifier:
22202811
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 401; Journal Issue: 2; Other Information: Copyright (c) 2010 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACTIN; IN VITRO; MASS SPECTROSCOPY; MONKEYS; RATS; SPERMATOGENESIS; SPERMATOZOA; VITAMIN A

Citation Formats

Zuo, Yan, Gao, Jing, Yeung, William S.B., Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Lee, Kai-Fai, and Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam. The testis-specific VAD1.3/AEP1 interacts with {beta}-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence. United States: N. p., 2010. Web. doi:10.1016/J.BBRC.2010.09.049.
Zuo, Yan, Gao, Jing, Yeung, William S.B., Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Lee, Kai-Fai, & Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam. The testis-specific VAD1.3/AEP1 interacts with {beta}-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence. United States. https://doi.org/10.1016/J.BBRC.2010.09.049
Zuo, Yan, Gao, Jing, Yeung, William S.B., Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Lee, Kai-Fai, and Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam. 2010. "The testis-specific VAD1.3/AEP1 interacts with {beta}-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence". United States. https://doi.org/10.1016/J.BBRC.2010.09.049.
@article{osti_22202811,
title = {The testis-specific VAD1.3/AEP1 interacts with {beta}-actin and syntaxin 1 and directs peri-nuclear/Golgi expression with bipartite nucleus localization (BNL) sequence},
author = {Zuo, Yan and Gao, Jing and Yeung, William S.B. and Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam and Lee, Kai-Fai and Centre for Reproduction, Development and Growth, Hong Kong Jockey Club Clinical Research Centre, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam},
abstractNote = {Research highlights: {yields} VAD1.3 interacts {beta}-actin and syntaxin 1. {yields} VAD1.3 colocalizes {beta}-actin in spermatids. {yields} The bipartite nucleus localization (BNL) signal is important for peri-nuclear/Golgi expression in transfected cells. {yields} The C-terminal region of VAD1.3 direct nuclei localization. -- 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 {beta}-actin proteins in vitro. Immunogold electron microscopic study localized VAD1.3 immunoreactivity to the acrosome membranes and matrix, and colocalized it with the {beta}-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 {beta}-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 {beta}-actin for acrosome formation in spermatogenesis.},
doi = {10.1016/J.BBRC.2010.09.049},
url = {https://www.osti.gov/biblio/22202811}, journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 2,
volume = 401,
place = {United States},
year = {Fri Oct 15 00:00:00 EDT 2010},
month = {Fri Oct 15 00:00:00 EDT 2010}
}