PMID- 10625686
OWN - NLM
STAT- MEDLINE
DCOM- 20000218
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 2
DP  - 2000 Jan 14
TI  - Role of cyclin G-associated kinase in uncoating clathrin-coated vesicles from
      non-neuronal cells.
PG  - 1365-70
AB  - Auxilin is a brain-specific DnaJ homolog that is required for Hsc70 to dissociate
      clathrin from bovine brain clathrin-coated vesicles. However, Hsc70 is also
      involved in uncoating clathrin-coated vesicles formed at the plasma membrane of
      non-neuronal cells suggesting that an auxilin homolog may be required for
      uncoating in these cells. One candidate is cyclin G-associated kinase (GAK), a
      150-kDa protein expressed ubiquitously in various tissues. GAK has a C-terminal
      domain with high sequence similarity to auxilin; like auxilin this C-terminal
      domain consists of three subdomains, an N-terminal tensin-like domain, a
      clathrin-binding domain, and a C-terminal J-domain. Western blot analysis shows
      that GAK is present in rat liver, bovine testes, and bovine brain clathrin-coated
      vesicles. More importantly, liver clathrin-coated vesicles, which contain GAK but
      not auxilin, are uncoated by Hsc70, suggesting that GAK acts as an auxilin
      homolog in non-neuronal cells. In support of this view, the clathrin-binding
      domain of GAK alone induces clathrin polymerization into baskets and the combined
      clathrin-binding domain and J-domain of GAK supports uncoating of AP180-clathrin 
      baskets by Hsc70 at pH 7 and induces Hsc70 binding to clathrin baskets at pH 6.
      Immunolocalization studies suggest that GAK is a cytosolic protein that is
      concentrated in the perinuclear region; it appears to be highly associated with
      the trans-Golgi where the budding of clathrin-coated vesicles occurs. We propose 
      that GAK is a required cofactor for the uncoating of clathrin-coated vesicles by 
      Hsc70 in non-neuronal cells.
FAU - Greener, T
AU  - Greener T
AD  - Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda,
      Maryland 20892-0301, USA.
FAU - Zhao, X
AU  - Zhao X
FAU - Nojima, H
AU  - Nojima H
FAU - Eisenberg, E
AU  - Eisenberg E
FAU - Greene, L E
AU  - Greene LE
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Adaptor Proteins, Vesicular Transport)
RN  - 0 (Clathrin)
RN  - 0 (Cyclins)
RN  - 0 (Gak protein, rat)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Monomeric Clathrin Assembly Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (clathrin assembly protein AP180)
RN  - EC 2.7.11.1 (GAK protein, human)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
SB  - IM
MH  - Adaptor Proteins, Vesicular Transport
MH  - Animals
MH  - Binding Sites
MH  - Brain/enzymology
MH  - Cattle
MH  - Clathrin/*metabolism/ultrastructure
MH  - Coated Pits, Cell-Membrane/*metabolism
MH  - Cyclins/*metabolism
MH  - Golgi Apparatus/metabolism/ultrastructure
MH  - HeLa Cells
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Kinetics
MH  - Liver/enzymology
MH  - Male
MH  - *Monomeric Clathrin Assembly Proteins
MH  - Nerve Tissue Proteins/metabolism
MH  - Phosphoproteins/metabolism
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Rats
MH  - Recombinant Proteins/metabolism
MH  - Testis/enzymology
MH  - Transfection
EDAT- 2000/01/08 09:00
MHDA- 2000/02/26 09:00
CRDT- 2000/01/08 09:00
PHST- 2000/01/08 09:00 [pubmed]
PHST- 2000/02/26 09:00 [medline]
PHST- 2000/01/08 09:00 [entrez]
AID - 10.1074/jbc.275.2.1365 [doi]
PST - ppublish
SO  - J Biol Chem. 2000 Jan 14;275(2):1365-70. doi: 10.1074/jbc.275.2.1365.