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doi: 10.1242/10.1242/jcs.00123
Research Article |
1 Institute of Molecular Medicine, National Taiwan University Hospital, College
of Medicine, National Taiwan University, Taipei, Taiwan, Republic of
China
2 Institute of Pathology, National Taiwan University Hospital, College of
Medicine, National Taiwan University, Taipei, Taiwan, Republic of China
* Author for correspondence (e-mail: fangjen{at}ha.mc.ntu.edu.tw)
Accepted 23 August 2002
ARF-like proteins (ARLs) are distinct group of members of the ARF family of
Ras-related GTPases. Although ARLs are very similar in primary structure to
ARFs, their functions remain unclear. We cloned mouse (m) and human (h) ARL5
cDNAs to characterize the protein products and their molecular properties.
mARL5 mRNA was more abundant in liver than in other adult tissues tested.
mARL5, similar to mARL4, was developmentally regulated and localized to
nuclei. hARL5 interacted with importin-
through its C-terminal
bipartite nuclear localization signal. When expressed in COS-7 cells, mutant
hARL5(T35N), which is predicted to be GDP bound, was concentrated in nucleoli.
The N-terminus of hARL5, like that of ARF, was myristoylated. Yeast two-hybrid
screening and in vitro protein-interaction assays showed that hARL5(Q80L),
predicted to be GTP bound, interacted with heterochromatin protein 1
(HP1
), which is known to be associated with telomeres as well as with
heterochromatin, and acted as a transcriptional suppressor in mammalian cells.
The interaction was reproduced in COS cells, where hARL5(Q80L) was
co-immunoprecipitated with HP1
. hARL5 interaction with HP1
was
dependent on the nucleotide bound, and required the MIR-like motif. Moreover,
hARL5(Q80L), but not hARL5 lacking the MIR-like motif, was partly co-localized
with overexpressed HP1
. Our findings suggest that developmentally
regulated ARL5, with its distinctive nuclear/nucleolar localization and
interaction with HP1
, may play a role(s) in nuclear dynamics and/or
signaling cascades during embryonic development.
Key words: ADP-ribosylation factors, Myristoylation, Heterochromatin, Importin-
, HP1
, Embryonic development
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