PMID- 21536677
OWN - NLM
STAT- MEDLINE
DCOM- 20110909
LR  - 20240320
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 286
IP  - 26
DP  - 2011 Jul 1
TI  - In vitro motility of liver connexin vesicles along microtubules utilizes kinesin 
      motors.
PG  - 22875-85
LID - 10.1074/jbc.M111.219709 [doi]
AB  - Trafficking of the proteins that form gap junctions (connexins) from the site of 
      synthesis to the junctional domain appears to require cytoskeletal delivery 
      mechanisms. Although many cell types exhibit specific delivery of connexins to 
      polarized cell sites, such as connexin32 (Cx32) gap junctions specifically 
      localized to basolateral membrane domains of hepatocytes, the precise roles of 
      actin- and tubulin-based systems remain unclear. We have observed fluorescently 
      tagged Cx32 trafficking linearly at speeds averaging 0.25 mum/s in a polarized 
      hepatocyte cell line (WIF-B9), which is abolished by 50 muM of the 
      microtubule-disrupting agent nocodazole. To explore the involvement of 
      cytoskeletal components in the delivery of connexins, we have used a preparation 
      of isolated Cx32-containing vesicles from rat hepatocytes and assayed their 
      ATP-driven motility along stabilized rhodamine-labeled microtubules in vitro. 
      These assays revealed the presence of Cx32 and kinesin motor proteins in the same 
      vesicles. The addition of 50 muM ATP stimulated vesicle motility along linear 
      microtubule tracks with velocities of 0.4-0.5 mum/s, which was inhibited with 1 mM 
      of the kinesin inhibitor AMP-PNP (adenylyl-imidodiphosphate) and by anti-kinesin 
      antibody but only minimally affected by 5 muM vanadate, a dynein inhibitor, or by 
      anti-dynein antibody. These studies provide evidence that Cx32 can be transported 
      intracellularly along microtubules and presumably to junctional domains in cells 
      and highlight an important role of kinesin motor proteins in 
      microtubule-dependent motility of Cx32.
FAU - Fort, Alfredo G
AU  - Fort AG
AD  - Dominick P. Purpura Department of Neuroscience, Albert Einstein College of 
      Medicine, Bronx, New York 10461, USA.
FAU - Murray, John W
AU  - Murray JW
FAU - Dandachi, Nadine
AU  - Dandachi N
FAU - Davidson, Michael W
AU  - Davidson MW
FAU - Dermietzel, Rolf
AU  - Dermietzel R
FAU - Wolkoff, Allan W
AU  - Wolkoff AW
FAU - Spray, David C
AU  - Spray DC
LA  - eng
GR  - P01 DK041918/DK/NIDDK NIH HHS/United States
GR  - DK41918/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20110502
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Connexins)
RN  - 25612-73-1 (Adenylyl Imidodiphosphate)
RN  - 3WHH0066W5 (Vanadates)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 3.6.4.2 (Dyneins)
RN  - EC 3.6.4.4 (Kinesins)
SB  - IM
MH  - Adenosine Triphosphate/chemistry/genetics/metabolism
MH  - Adenylyl Imidodiphosphate/chemistry/genetics/metabolism
MH  - Animals
MH  - Cell Line, Tumor
MH  - Connexins/chemistry/genetics/*metabolism
MH  - Dyneins/chemistry/genetics/metabolism
MH  - Gap Junctions/chemistry/genetics/metabolism
MH  - Hepatocytes/chemistry/*metabolism
MH  - Humans
MH  - Kinesins/chemistry/genetics/*metabolism
MH  - Liver/chemistry/*metabolism
MH  - Microtubules/chemistry/genetics/*metabolism
MH  - Protein Transport/drug effects/physiology
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Vanadates/chemistry
MH  - Gap Junction beta-1 Protein
PMC - PMC3123055
EDAT- 2011/05/04 06:00
MHDA- 2011/09/10 06:00
PMCR- 2012/07/01
CRDT- 2011/05/04 06:00
PHST- 2011/05/04 06:00 [entrez]
PHST- 2011/05/04 06:00 [pubmed]
PHST- 2011/09/10 06:00 [medline]
PHST- 2012/07/01 00:00 [pmc-release]
AID - S0021-9258(19)48825-X [pii]
AID - M111.219709 [pii]
AID - 10.1074/jbc.M111.219709 [doi]
PST - ppublish
SO  - J Biol Chem. 2011 Jul 1;286(26):22875-85. doi: 10.1074/jbc.M111.219709. Epub 2011 
      May 2.