Evidence that Golgi structure depends on a p115 activity that is independent of the vesicle tether components giantin and GM130

doi:10.1083/jcb.200105005

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Published online 8 October 2001. doi:10.1083/jcb.200105005

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© The Rockefeller University Press, 0021-9525/2001/10/227 $5.00
The Journal of Cell Biology, Volume 155, Number 2, October 15, 2001 227-238

Article

Evidence that Golgi structure depends on a p115 activity that is independent of the vesicle tether components giantin and GM130

Manojkumar A. Puthenveedu and Adam D. Linstedt

Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213

Address correspondence to Adam D. Linstedt, Dept. of Biological Sciences, Carnegie Mellon University, 4400 5th Ave., Pittsburgh, PA 15213. Tel.: (412) 268-1249. Fax: (412) 268-7129. E-mail: linstedt{at}andrew.cmu.edu

Inhibition of the putative coatomer protein I (COPI) vesicletethering complex, giantin–p115–GM130, may contributeto mitotic Golgi breakdown. However, neither this, nor the roleof the giantin–p115–GM130 complex in the maintenanceof Golgi structure has been demonstrated in vivo. Therefore,we generated antibodies directed against the mapped bindingsites in each protein of the complex and injected these intomammalian tissue culture cells. Surprisingly, the injected anti-p115and antigiantin antibodies caused proteasome-mediated degradationof the corresponding antigens. Reduction of p115 levels belowdetection led to COPI-dependent Golgi fragmentation and apparentaccumulation of Golgi-derived vesicles. In contrast, neitherreduction of giantin below detectable levels, nor inhibitionof p115 binding to GM130, had any detectable effect on Golgistructure or Golgi reassembly after cell division or brefeldinA washout. These observations indicate that inhibition of p115can induce a mitotic-like Golgi disassembly, but its essentialrole in Golgi structure is independent of its Golgi-localizedbinding partners giantin and GM130.

Key Words: Golgi structure; tether; docking; mitosis; COPI

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