Fission yeast translation initiation factor 3 subunit eIF3h is not essential for global translation initiation, but deletion of eif3h+ affects spore formation.
The fission yeast Schizosaccharomyces pombe homologue of the p40/eIF3h subunit of mammalian translation initiation factor eIF3 has been characterized in this study. We show that this protein physically associates with the 40S ribosomal particles as a constituent of the multimeric eIF3 protein complex, which consists of all five known eIF3 core subunits (eIF3a, eIF3b, eIF3c, eIF3g and eIF3i) as well as the five non-core subunits (eIF3d, eIF3e, eIF3f, eIF3h and eIF3m) that constitute an eIF3 holocomplex in fission yeast. However, affinity purification of eIF3 from fission yeast cells expressing TAP-tagged eIF3h suggests the presence of distinct forms of eIF3 that differ in their composition of the non-core subunits. Further characterization of eIF3h shows that strains lacking eif3h(+) (eif3hDelta) are viable and show no gross defects, either in vegetative growth or in the rate of in vivo protein synthesis. Polysome profile analysis shows no apparent defects in translation initiation. Furthermore, deletion of eif3h(+) does not affect the ability of the other eIF3 subunits to remain associated with one another in a tight protein complex similar to the situation in wild-type cells. Additionally, we show that human eIF3h can functionally substitute fission yeast eIF3h in complementing in vivo a genetic deletion of eif3h(+). Interestingly, mutant eif3hDelta cells show several prominent phenotypic properties. They are hypersensitive to caffeine and highly defective in meiosis, producing either no spores or incomplete tetrads with a very high frequency. The implications of these results in relation to the functions of eIF3h in Sz. pombe are discussed.
Ray
Anirban
A
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine of Yeshiva University, Bronx, New York, NY 10461, USA.
Bandyopadhyay
Amitabha
A
Matsumoto
Tomohiro
T
Deng
Haiteng
H
Maitra
Umadas
U
D000935:Antifungal Agents / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology; D039621:Eukaryotic Initiation Factor-3 / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D050296:Microbial Viability; D011485:Protein Binding; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000737:chemistry; D012568:Schizosaccharomyces / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D013172:Spores, Fungal / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*
D000935:Antifungal Agents; D039621:Eukaryotic Initiation Factor-3; D002110:Caffeine