Publication: How can mammalian Rab small
GTPases be comprehensively analyzed?:
Development of new tools to comprehensively
analyze mammalian Rabs in membrane traffic
Authors
Fukuda, Mitsunori
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Small GTPase Rabs constitute the largest
family of membrane trafficking proteins that are
conserved in all eukaryotic cells. The number of
different Rab isoforms in multicellular organisms is
usually greater than that in unicellular organisms (e.g.,
approximately 60 different Rabs in each species of
mammals investigated versus approximately 10 Rabs in
yeasts). The expansion of Rab isoforms in mammals is
often regarded as due to the acquisition of specialized
membrane trafficking events in the specialized cell types
of higher eukaryotes. However, because of their large
numbers the precise function of most mammalian Rab
isoforms is still unknown. The recent development of
new tools for comprehensive analysis (e.g., Rab panels)
has paved the way for systematic investigation of the
involvement of mammalian Rab isoforms in specialized
membrane trafficking events. The tools include
collections of enhanced green fluorescent protein
(EGFP)-tagged mouse and human Rabs, FLAG-tagged
Rabs, glutathione S-transferase (GST)-tagged Rabs,
Gal4-binding domain (GBD)-tagged Rabs, Tre-
2/Bub2/Cdc16 (TBC) domain-containing Rab-GTPaseactivating
proteins (GAPs), and small interfering RNAs.
EGFP-Rabs are used to screen for Rabs that are
localized on specific organelles and regulate their
transport, and GST-Rabs and GBD-Rabs are used to
screen for novel Rab effectors by GST pull-down assays
and yeast two-hybrid assays, respectively. Combined use
of these tools now makes it possible to efficiently
determine the function of mammalian Rab isoforms in
membrane traffic. This article reviews the development
of new tools for systematic analysis of Rab proteins and
their application to Rab-mediated membrane traffic.
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