Ohno K, Hirose F, Inoue Y H, Takisawa H, Mimura S, Hashimoto Y, Kiyono T, Nishida Y, Matsukage A
Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya 464-8681, Japan.
Gene. 1998 Sep 14;217(1-2):177-85. doi: 10.1016/s0378-1119(98)00358-8.
cDNAs encoding three Drosophila melanogaster MCM proteins, DmMCM3, DmMCM6 and DmMCM7, candidates of DNA replication-licensing factors, were cloned and sequenced. The deduced amino-acid sequences displayed 60, 59 and 68% identities with the respective Xenopus laevis homologues, XMCM3, XMCM6 and XMCM7. Six members of the D. melanogaster MCM family were found to share 31-36% identities in their amino-acid sequences, and to possess the five common domains carrying conserved amino-acid sequences as reported with X. laevis MCM proteins. DmMCM3, DmMCM6 and DmMCM7 genes were mapped to the 4F region on the X chromosome, the 6B region on the X chromosome and the 66E region on the third chromosome, respectively, by in situ hybridization. Contents of their mRNAs were proved to be high in unfertilized eggs and early embryos (0-4h after fertilization), then decrease gradually by the 12h time point, with only low levels detected at later stages of development except in adult females. This fluctuation pattern is similar to those of genes for proteins involved in DNA replication, such as DNA polymerase alpha and proliferating cell nuclear antigen, suggesting that expression of DmMCM genes is under the regulatory mechanism which regulates expression of other genes involved in DNA replication.
编码三种果蝇MCM蛋白(DmMCM3、DmMCM6和DmMCM7)的cDNA被克隆并测序,这三种蛋白是DNA复制许可因子的候选蛋白。推导的氨基酸序列与相应的非洲爪蟾同源物XMCM3、XMCM6和XMCM7分别具有60%、59%和68%的同一性。发现果蝇MCM家族的六个成员在氨基酸序列上具有31%-36%的同一性,并拥有五个带有保守氨基酸序列的共同结构域,这与非洲爪蟾MCM蛋白的报道一致。通过原位杂交,DmMCM3、DmMCM6和DmMCM7基因分别被定位到X染色体的4F区域、X染色体的6B区域和第三条染色体的66E区域。结果证明,它们的mRNA含量在未受精卵和早期胚胎(受精后0-4小时)中较高,然后在12小时时间点逐渐下降,除成年雌性外,在发育后期仅检测到低水平。这种波动模式与参与DNA复制的蛋白质基因(如DNA聚合酶α和增殖细胞核抗原)的波动模式相似,表明DmMCM基因的表达受调控其他参与DNA复制基因表达的调控机制的控制。