Subcellular localization and membrane association of SARS-CoV 3a protein.
SARS-CoV 3a protein was a unique protein of SARS coronavirus (SARS-CoV), which was identified in SARS-CoV infected cells and SARS patients' specimen. Recent studies revealed that 3a could interact specifically with many SARS-CoV structural proteins, such as M, E and S protein. Expressed 3a protein was reported to localize to Golgi complex in SARS-CoV infected cells. In this study, it was shown that 3a protein was mainly located in Golgi apparatus with different tags at N- or C-terminus. The localization pattern was similar in different transfected cells. With the assay of truncated 3a protein, it was shown that 3a might contain three transmembrane regions, and the second or third region was properly responsible for Golgi localization. By ultra-centrifugation experiment with different extraction buffers, it was confirmed that 3a was an integral membrane protein and embedded in the phospholipid bilayer. Immunofluorescence assay indicated that 3a was co-localized with M protein in Golgi complex in co-transfected cells. These results provide a new insight for further study of the 3a protein on the pathogenesis of SARS-CoV.
Yuan
Xiaoling
X
Department of Pathophysiology, Beijing Institute of Radiation Medicine, No. 27 Taiping Road, Beijing, 100850, China.
Li
Jianyong
J
Shan
Yajun
Y
Yang
Zhen
Z
Zhao
Zhenhu
Z
Chen
Bo
B
Yao
Zhenyu
Z
Dong
Bo
B
Wang
Shengqi
S
Chen
Jiapei
J
Cong
Yuwen
Y
D000818:Animals; D019556:COS Cells; D002458:Cell Fractionation; D002460:Cell Line; D002522:Chlorocebus aethiops; D000086862:Coronavirus M Proteins; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000737:chemistry; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis; D017434:Protein Structure, Tertiary; D045473:Severe acute respiratory syndrome-related coronavirus; D013347:Subcellular Fractions / Q000737:chemistry*; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins / Q000032:analysis; D014764:Viral Proteins / Q000032:analysis* / Q000737:chemistry; D000086082:Viroporin Proteins
C487105:3a protein, SARS-CoV; D000086862:Coronavirus M Proteins; D008051:Lipid Bilayers; C000706237:M protein, SARS-CoV; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D000086082:Viroporin Proteins