Kaneko S, Miyazaki Y, Yasuda T, Shishido K
Department of Life Science, Faculty of Bioscience, Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
Gene. 1998 May 12;211(2):259-66. doi: 10.1016/s0378-1119(98)00099-7.
Sequence analysis of the downstream region of the basidiomycete Lentinus edodes priB gene encoding a protein with a 'Zn(II)2Cys6 zinc cluster' DNA-binding motif (Endo, H., Kajiwara, S., Tunoka, O., Shishido, K., 1994. A novel cDNA, priBc, encoding a protein with a Zn(II)2Cys6 zinc cluster DNA-binding motif, derived from the basidiomycete Lentinus edodes. Gene 139, 117-121) suggested the presence of a Saccharomyces cerevisiae URA6 gene homologue encoding UMP kinase. We isolated a corresponding cDNA from a mature fruiting-body cDNA library of L. edodes. The nucleotide sequence of this was determined and compared with that of the genomic DNA, revealing that the URA6 gene homologue encodes 227 amino acids (aa) and is interrupted by four small introns. The deduced aa sequence showed an overall identity of 51.1% to that of the S. cerevisiae URA6 gene product. The URA6 homologue protein produced in Escherichia coli using the glutathione S-transferase gene fusion system was found to catalyze the phosphoryl transfer from ATP to UMP and CMP efficiently and also to AMP and dCMP with lower efficiencies. Thus, the URA6 gene homologue was designated uck1 and its product UMP-CMP kinase. Northern-blot analysis showed that the uck1 is actively transcribed in the gill tissue of mature fruiting bodies of L. edodes, implying that uck1 may play a role during the formation of basidiospores occurs in the gill tissue.
对担子菌香菇priB基因下游区域进行序列分析，该基因编码一种具有“Zn(II)2Cys6锌簇”DNA结合基序的蛋白质（远藤浩、梶原幸、富野雄、宍戸和夫，1994年。一种新的cDNA，priBc，编码一种具有Zn(II)2Cys6锌簇DNA结合基序的蛋白质，源自担子菌香菇。基因139，117 - 121），结果表明存在一个编码UMP激酶的酿酒酵母URA6基因同源物。我们从香菇成熟子实体cDNA文库中分离出了一个相应的cDNA。测定了其核苷酸序列，并与基因组DNA序列进行比较，结果显示URA6基因同源物编码227个氨基酸，且被四个小内含子打断。推导的氨基酸序列与酿酒酵母URA6基因产物的总体一致性为51.1%。利用谷胱甘肽S - 转移酶基因融合系统在大肠杆菌中产生的URA6同源蛋白被发现能够高效地催化从ATP到UMP和CMP的磷酰基转移，对AMP和dCMP的催化效率较低。因此，URA6基因同源物被命名为uck1，其产物为UMP - CMP激酶。Northern杂交分析表明，uck1在香菇成熟子实体的菌褶组织中活跃转录，这意味着uck1可能在菌褶组织中担孢子形成过程中发挥作用。
