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MBC in Press, published online ahead of print December 25, 2002
Mol. Biol. Cell 10.1091/mbc.E02-08-0504

A more recent version of this article appeared on March 1, 2003
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Submitted on August 15, 2002
Revised on October 18, 2002
Accepted on November 18, 2002

The Fission Yeast spo14+ Gene Encoding a Functional Homologue of Budding Yeast Sec12 Is Required for the Development of Forespore Membranes

Michiko Nakamura-Kubo1, Taro Nakamura1, Aiko Hirata2, and Chikashi Shimoda1*

1 Department of Biology, Graduate School of Science, Osaka City University, Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
2 Department of Integrated Biosciences, Graduate School of Frontier Science, University of Tokyo, Kashiwa, Chiba 277-8562, Japan

* Corresponding author. E-mail address: shimoda{at}sci.osaka-cu.ac.jp.

The Schizosaccharomyces pombe spo14-B221 mutant was originally isolated as a sporulation-deficient mutant. However, the spo14+ gene is essential for cell viability and growth. spo14+ is identical to the previously characterized stl1i+gene encoding a putative homologue of Saccharomyces cerevisiae Sec12, which is essential for protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. In the spo14 mutant cells, ER-like membranes were accumulated beneath the plasma membrane and the ER/Golgi shuttling protein Rer1 remained in the ER. Sec12 is a guanine nucleotide exchange factor for the Sar1 GTPase. Overproduction of psr1+ coding for an S. pombe Sar1 homologue suppressed both the sporulation defect of spo14-B221 and cold-sensitive growth of newly isolated spo14-6 and spo14-7 mutants. These results indicate that Spo14 is involved in early steps of the protein secretory pathway. The spo14-B221 allele carries a single nucleotide change in the branch point consensus of the fifth intron, which reduces the abundance of the spo14 mRNA. During meiosis II, the forespore membrane was initiated near spindle pole bodies; however, subsequent extension of the membrane was arrested before its closure into a sac. We conclude that Spo14 is responsible for the assembly of the forespore membrane by supplying membrane vesicles.




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