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J. Biol. Chem., Vol. 276, Issue 34, 32056-32063, August 24, 2001
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,
, and
From the Biotechnology Centre of Oslo, University of Oslo, P.O. Box
1125 Blindern, N-0317 Oslo, Norway
A cDNA encoding a novel member of the
helicase family, MDDX28, has been cloned from a human testis
library. This apparently intronless gene was transcribed in all
tissues studied. MDDX28 encodes a protein of 540 amino acids, with
~30% homology to other helicases over the core region, containing
all the conserved DEAD-box helicase motifs. No homologue is known.
MDDX28 has RNA and Mg2+-dependent ATPase
activity. Subcellular localization studies of MDDX28 using
oligoclonal antibodies raised against the protein as well as its
enhanced green fluorescence protein (EGFP) demonstrated that the
protein is localized in the mitochondria and the nucleus. To our
knowledge, MDDX28 is the first member of the RNA helicase described
with this dual location. The nuclear localization of MDDX28
depended on active RNA polymerase II transcription, suggesting that the
protein could be transported to and from the nucleus. This was
confirmed further in an interspecies heterokaryon assay, in which
MDDX28 was seen to translocate to the nucleus and mitochondria. The
mitochondrial uptake of the MDDX28-EGFP-N1 fusion protein was
inhibited by carbonyl cyanide
p-(trichloromethoxy)phenylhydrazone. Our results indicate
that MDDX28 can be transported between the mitochondria and the nucleus.
The sequence data reported in this article have a GenBank Accession Number AF329821. This sequence is also covered by AC004531, AK002144 and XM_007885. The corresponding murine DNA sequence was submitted February 1, 2001, and has the Accession Number BAB26907.
Research Fellow of the Norwegian Cancer Society.
§
Research Fellow of the Norwegian Cancer Society.
¶
Present address: Institute for Nutrition Research, University
of Oslo, Oslo, Norway.
Postdoctoral Fellow of the Norwegian Cancer Society.
**
To whom correspondence should be addressed. Tel.: 47-22-840531;
Fax: 47-22-840501; E-mail: hans.prydz@biotek.uio.no.
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