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PMID: 17093494 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cargo selection by specific kinesin light chain 1 isoforms.

The EMBO journal ·Vol. 25 ·No. 23 ·2006-11-29 ·Pages 5457-68

Woźniak MJ, Allan VJ

Abstract

Kinesin-1 drives the movement of diverse cargoes, and it has been proposed that specific kinesin light chain (KLC) isoforms target kinesin-1 to these different structures. Here, we test this hypothesis using two in vitro motility assays, which reconstitute the movement of rough endoplasmic reticulum (RER) and vesicles present in a Golgi membrane fraction. We generated GST-tagged fusion proteins of KLC1B and KLC1D that included the tetratricopeptide repeat domain and the variable C-terminus. We find that preincubation of RER with KLC1B inhibits RER motility, whereas KLC1D does not. In contrast, Golgi fraction vesicle movement is inhibited by KLC1D but not KLC1B reagents. Both RER and vesicle movement is inhibited by preincubation with the GST-tagged C-terminal domain of ubiquitous kinesin heavy chain (uKHC), which binds to the N-terminal domain of uKHC and alters its interaction with microtubules. We propose that although the TRR domains are required for cargo binding, it is the variable C-terminal region of KLCs that are vital for targeting kinesin-1 to different cellular structures.

MeSH Terms
Alternative Splicing Amino Acid Sequence Animals Biological Assay Cytoplasmic Vesicles/drug effects Endoplasmic Reticulum/drug effects Golgi Apparatus/chemistry,drug effects,ultrastructure Intracellular Membranes/drug effects Kinesins Microtubule-Associated Proteins/analysis,genetics,pharmacology Molecular Sequence Data Protein Isoforms/analysis,genetics,pharmacology Rats Recombinant Fusion Proteins/pharmacology
Chemicals
Klc1 protein, rat Microtubule-Associated Proteins Protein Isoforms Recombinant Fusion Proteins Kinesins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Woźniak Marcin J
Faculty of Life Sciences, University of Manchester, Manchester, UK.
Allan Victoria J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7601427
Published
2006-11-29
Epub
2006-00-09
Pages
5457-68
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1679764
Subset
IM
Grants
Wellcome Trust · 078825/Z/05/Z · United Kingdom
External Links
PubMed source DOI: 10.1038/sj.emboj.7601427 PMC Full Text View journal details
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