Nakazawa Takehito, Kaneko Shinya, Miyazaki Yasumasa, Jojima Toru, Yamazaki Takashi, Katsukawa Shiho, Shishido Kazuo
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
Fungal Genet Biol. 2008 Jun;45(6):818-28. doi: 10.1016/j.fgb.2008.02.007. Epub 2008 Feb 29.
We isolated a target gene for the Lentinula edodes putative transcription factor Le.CDC5 that contains a c-Myb-type DNA-binding domain. The gene, termed ctg1, encodes a novel protein (159 amino acid residues) with a leucine zipper-like sequence and contains a 7-bp Le.CDC5-binding sequence, 5'GCAATCT3', in its transcribed region downstream of the start codon. Chromatin immunoprecipitation analysis strongly suggested that intracellular Le.CDC5 binds to this 7-bp sequence on L. edodes chromatin. Binding was most efficient on chromatin from the stipes of mature fruiting bodies. Two Le.CDC5-interaction partners were identified in L. edodes and named CIPA and CIPB. The CIPB protein (127 amino acid residues) binds to a 6-bp sequence with the consensus sequence 5'CAACAC/T/G3'. The ctg1 gene contains nine 6-bp consensus (or consensus-like) sequences, six are in the 5'-upstream region and three in the transcribed region downstream of the start codon. At least two each of the upstream and downstream sequences appear to bind CIPB in vitro. We suggest that Le.CDC5 and CIPB can cooperatively regulate the expression of ctg1.
我们分离出了香菇假定转录因子Le.CDC5的一个靶基因,该基因含有一个c-Myb型DNA结合结构域。这个名为ctg1的基因编码一种具有亮氨酸拉链样序列的新型蛋白质(159个氨基酸残基),并且在其起始密码子下游的转录区域含有一个7碱基对的Le.CDC5结合序列5'GCAATCT3'。染色质免疫沉淀分析强烈表明,细胞内的Le.CDC5与香菇染色质上的这个7碱基对序列结合。在成熟子实体菌柄的染色质上结合效率最高。在香菇中鉴定出了两个Le.CDC5相互作用伙伴,分别命名为CIPA和CIPB。CIPB蛋白(127个氨基酸残基)与一个共有序列为5'CAACAC/T/G3'的6碱基对序列结合。ctg1基因包含九个6碱基对的共有(或类似共有)序列,六个在5'上游区域,三个在起始密码子下游的转录区域。上游和下游序列中至少各有两个在体外似乎能与CIPB结合。我们认为Le.CDC5和CIPB可以协同调节ctg1的表达。