Sista H, Wechser M A, Bowman B J
Department of Biology, Sinsheimer Labs, University of California, Santa Cruz 95064.
Mol Gen Genet. 1994 Apr;243(1):82-90. doi: 10.1007/BF00283879.
We have isolated the proteolipid subunit from the vacuolar ATPase of Neurospora crassa, using ion-exchange chromatography. We have also isolated several cDNA clones and the corresponding genomic DNA that encodes this subunit. The derived protein sequence indicates that the polypeptide is composed of 161 amino acid residues with an M(r) of 16,328 kDa. The gene encoding the proteolipid, named vma-3, is unusual in several respects. It contains four introns and, unlike other fungal genes, has non-coding regions that are as large as the coding regions. The 3' untranslated regions of the cDNAs were quite heterogeneous, with polyadenylation sites more than 300 bp apart. Analysis of the mRNA indicates that two size classes of transcripts are produced, differing in the length of the 3' untranslated region. Mapping of the vma-3 gene showed that it is closely linked, but not adjacent to, vma-1, the gene encoding the 67 kDa subunit of the vacuolar ATPase. This raises the possibility that in N. crassa some of the vacuolar ATPase genes may be clustered.
我们利用离子交换色谱法从粗糙脉孢菌的液泡ATP酶中分离出了蛋白脂质亚基。我们还分离出了几个cDNA克隆以及编码该亚基的相应基因组DNA。推导的蛋白质序列表明，该多肽由161个氨基酸残基组成，分子量为16,328 kDa。编码该蛋白脂质的基因名为vma - 3，在几个方面都很不寻常。它包含四个内含子，并且与其他真菌基因不同，其非编码区与编码区一样大。cDNA的3'非翻译区非常不均一，聚腺苷酸化位点相隔超过300 bp。对mRNA的分析表明产生了两种大小类别的转录本，其3'非翻译区的长度不同。vma - 3基因的定位显示它与液泡ATP酶67 kDa亚基的编码基因vma - 1紧密连锁，但不相邻。这增加了在粗糙脉孢菌中一些液泡ATP酶基因可能成簇的可能性。
