Elf1p, a member of the ABC class of ATPases, functions as a mRNA export factor in Schizosacchromyces pombe.
Rae1p and Mex67p/Tap are conserved mRNA export factors. We have used synthetic lethal genetic screens in Schizosaccharomyces pombe to identify mutations in genes that are functionally linked to rae1 and mex67 in mRNA export. From these screens, we have isolated mutations in a putative S. pombe homologue of the Candida albicans elf1 gene. The elf1 of S. pombe is not an essential gene. When elf1 mutations are combined with rae1-167 mutation, growth and mRNA export is inhibited in the double mutants. This inhibition can be suppressed by the multicopy expression of mex67 suggesting that Mex67p can substitute for the loss of Elf1p function. Elf1p is a non-membrane member of the ATP-binding cassette (ABC) class of ATPase and the GFP-Elf1p fusion localizes to the cytoplasm. Elf1p, expressed and purified from Escherichia coli, binds and hydrolyzes ATP. A mutant Elf1p that carries a glycine to aspartic acid (G731D) mutation within the Walker A domain of the second ATP site retains the ATP binding but loses its ATPase activity in vitro. This mutant protein no longer functions in mRNA export. Taken together, our results show that Elf1p functions as a mRNA export factor along with Rae1p and Mex67p in S. pombe.
Basic Research Laboratory, Center for Cancer Research, NCI/National Institutes of Health, Bldg. 37/Rm. 6138B, 9000 Rockville Pike, Bethesda, MD 20892, USA.
Libor
L
Kozak
Ganesh
G
Gopal
Jin Ho
JH
Yoon
Zuben E
ZE
Sauna
Suresh V
SV
Ambudkar
Anjan G
AG
Thakurta
Ravi
R
Dhar
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D021581:Active Transport, Cell Nucleus; D000251:Adenosine Triphosphatases / Q000502:physiology*; D000595:Amino Acid Sequence; D001692:Biological Transport; D005656:Fungal Proteins / Q000502:physiology*; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000378:metabolism; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D012333:RNA, Messenger; D000251:Adenosine Triphosphatases