Increase in beta1-6 GlcNAc branching caused by N-acetylglucosaminyltransferase V directs integrin beta1 stability in human hepatocellular carcinoma cell line SMMC-7721.
In this study, an enzymatic inactive mutant of GnT-V (delta cGnT-V) was constructed and transfected in SMMC 7721 cell line. Integrin beta1 in delta cGnT-V transfectants (delta c-7721) showed attenuation of the number of beta1-6 GlcNAc branching, whereas those in wtGnT-V transfectants (wt-7721) presented a beta1-6 GlcNAc-rich pattern. High integrin beta1 expression was observed in wt-7721 compared with mock cells (7721 cell transfected with the vector pcDNA3), while transfection of delta cGnT-V decreased the integrin beta1 expression, despite of no significant changes on integrin beta1 mRNA level in these cell lines. Pulse-chase experiment showed that Integrin beta1 in delta c-7721 was prone to quick degradation and its half-life was less than 3 h, on the contrary, the alleviating degradation of beta1 subunit was observed in wt-7721 where the beta1 subunit half-life was about 16 h, meanwhile, the degradation rate of beta1 subunit in mock cells was in between, about 10 h. More effective in promoting cell migration toward fibronectin and invasion through Matrigel was observed in wt-7721 while this was almost suppressed in delta c-7721. Our results suggest that the addition of beta1-6 GlcNAc branching caused more fully glycosylated mature form on integrin beta1 and inhibited beta1 protein degradation. Glycosylation caused by GnT-V directs integrin beta1 stability and more delivery to plasma membrane, subsequently promotes Fn-based cell migration and invasion.
Wang
Liying
L
Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Liang
Yulong
Y
Li
Zengxia
Z
Cai
Xiumei
X
Zhang
Wen
W
Wu
Guoqiang
G
Jin
Jiawei
J
Fang
Zhengyu
Z
Yang
Yong
Y
Zha
Xiliang
X
D000117:Acetylglucosamine / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D005353:Fibronectins / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism*; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D011499:Protein Processing, Post-Translational
D005353:Fibronectins; D019012:Integrin beta1; D017351:N-Acetylglucosaminyltransferases; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase; D000117:Acetylglucosamine