GRIF-1 [GABAA (γ-aminobutyric acidA) receptor interacting factor-1] is a member of a coiled-coil family of proteins thought to function as adaptors in the anterograde trafficking of organelles utilizing the kinesin-1 motor proteins to synapses. To study in more detail the molecular interaction between GRIF-1 and the kinesin-1 family member KIF5C, fluorescent yellow- and fluorescent cyan-tagged GRIF-1, KIF5C, the KIF5C MD (motor domain) and the KIF5C NMD (non-motor domain) fusion proteins were generated. Each was characterized with respect to size and ability to co-associate by immunoprecipitation following expression in HEK-293 (human embryonic kidney 293) cells. Further, their distribution in transfected HEK-293 and transformed African green monkey kidney (COS-7) cells was analysed by confocal microscopy. The fluorescent GRIF-1 and KIF5C fusion proteins were all found to behave as wild-type. Double GRIF-1/KIF5C transfectants revealed co-localization. The GRIF-1/KIF5C and GRIF-1/KIF5C NMD double transfectants showed different subcellular distributions compared with single GRIF-1, KIF5C or KIF5C NMD transfections. These studies confirm the association between GRIF-1 and kinesin-1 NMDs. Fluorescence resonance energy transfer studies are ongoing to characterize this interaction in more detail.
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February 2006
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Conference Article|
January 20 2006
GRIF-1–kinesin-1 interactions: a confocal microscopy study
K. Pozo;
K. Pozo
1School of Pharmacy, University of London, 29/39 Brunswick Square, London WC1N 1AX, U.K.
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F.A. Stephenson
F.A. Stephenson
1
1School of Pharmacy, University of London, 29/39 Brunswick Square, London WC1N 1AX, U.K.
1 To whom correspondence should be addressed (email anne.stephenson@ulsop.ac.uk).
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Publisher: Portland Press Ltd
Received:
September 19 2005
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2006 The Biochemical Society
2006
Biochem Soc Trans (2006) 34 (1): 48–50.
Article history
Received:
September 19 2005
Citation
K. Pozo, F.A. Stephenson; GRIF-1–kinesin-1 interactions: a confocal microscopy study. Biochem Soc Trans 1 February 2006; 34 (1): 48–50. doi: https://doi.org/10.1042/BST0340048
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