Hodgetts R B, Clark W C, O'Keefe S L, Schouls M, Crossgrove K, Guild G M, von Kalm L
Department of Biological Sciences, University of Alberta, Edmonton, Canada.
Development. 1995 Nov;121(11):3913-22. doi: 10.1242/dev.121.11.3913.
The 2B5 early puff locus corresponds to the Broad-Complex BR-C) and encodes a family of transcription factors whose members are induced by the molting hormone ecdysone. Mutations in the br subcomplementation group substantially reduce the levels of Dopa decarboxylase (DDC) in the epidermis of mature third instar larvae but not in mature second instar organisms. Enzyme levels are normal in the central nervous system of the two mutants examined. The specificity of these effects suggests that a product of the BR-C locus mediates the rapid appearance of DDC in mature third instar larvae experiencing an elevated titer of ecdysone. The likely identity of this protein has been confirmed by pursuing the observation that the br28 allele caused by the insertion of a P element into the Z2 DNA-binding domain. Both the transcript and a protein carrying this domain are present in the epidermis and a BR-C recombinant protein carrying the Z2 finger binds to the first intron of the Ddc gene. Five binding sites have been identified within the intron by DNAase I footprinting and a core consensus sequence has been derived which shares some identity with the consensus binding site of the Z2 protein to the Sgs-4 regulatory region. Our demonstration that Ddc is a target of BR-C in the epidermis is the first direct evidence of a role for this early gene in a tissue other than the salivary glands. The data reinforce the idea that BR-C, which clearly mediates a salivary gland-specific response to ecdysone, may play a widespread role in the hormone's activation of gene cascades in other target tissues.
2B5早期膨大区位点对应于宽复合体(BR-C),并编码一类转录因子,其成员由蜕皮激素ecdysone诱导产生。br亚互补组中的突变显著降低了成熟三龄幼虫表皮中多巴脱羧酶(DDC)的水平,但在成熟二龄生物体中未降低。在所检测的两个突变体的中枢神经系统中,酶水平正常。这些效应的特异性表明,BR-C位点的一个产物介导了在蜕皮激素滴度升高的成熟三龄幼虫中DDC的快速出现。通过追踪观察到br28等位基因是由一个P元件插入Z2 DNA结合结构域引起的,证实了这种蛋白质的可能身份。携带该结构域的转录本和蛋白质均存在于表皮中,并且携带Z2指的BR-C重组蛋白与Ddc基因的第一个内含子结合。通过DNA酶I足迹法在该内含子内鉴定出了五个结合位点,并推导了一个核心共有序列,该序列与Z2蛋白与Sgs-4调控区的共有结合位点有一定的同源性。我们证明Ddc是表皮中BR-C的一个靶标,这是该早期基因在唾液腺以外的组织中发挥作用的第一个直接证据。这些数据强化了这样一种观点,即BR-C虽然明显介导了唾液腺对蜕皮激素的特异性反应,但可能在该激素激活其他靶组织中的基因级联反应中发挥广泛作用。