Parthasarthy Akhila, Gopinathan Karumathil P
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India.
FEBS J. 2005 Oct;272(20):5191-205. doi: 10.1111/j.1742-4658.2005.04877.x.
Members of a multigene family from the silkworm Bombyx mori have been classified based on their transcriptions in homologous nuclear extracts, into three groups of highly, moderately and poorly transcribed genes. Because all these gene copies have identical coding sequences and consequently identical promoter elements (the A and B boxes), the flanking sequences modulate their expression levels. Here we demonstrate the interaction of transcription factor TFIIIB with these genes and its role in regulating differential transcriptions. The binding of TFIIIB to the poorly transcribed gene -6,7 was less stable compared with binding of TFIIIB to the highly expressed copy, -1. The presence of a 5' upstream TATA sequence closer to the coding region in -6,7 suggested that the initial binding of TFIIIC to the A and B boxes sterically hindered anchoring of TFIIIB via direct interactions, leading to lower stability of TFIIIC-B-DNA complexes. Also, the multiple TATATAA sequences present in the flanking regions of this poorly transcribed gene successfully competed for TFIIIB reducing transcription. The transcription level could be enhanced to some extent by supplementation of TFIIIB but not by TATA box binding protein. The poor transcription of -6,7 was thus attributed both to the formation of a less stable transcription complex and the sequestration of TFIIIB. Availability of the transcription factor TFIIIB in excess could serve as a general mechanism to initiate transcription from all the individual members of the gene family as per the developmental needs within the tissue.
家蚕多基因家族的成员已根据其在同源核提取物中的转录情况,被分为高度转录、中度转录和低度转录的三组基因。由于所有这些基因拷贝都具有相同的编码序列,因此具有相同的启动子元件(A盒和B盒),侧翼序列调节它们的表达水平。在此,我们展示了转录因子TFIIIB与这些基因的相互作用及其在调节差异转录中的作用。与TFIIIB与高表达拷贝-1的结合相比,TFIIIB与低转录基因-6,7的结合不太稳定。-6,7中靠近编码区的5'上游TATA序列的存在表明,TFIIIC最初与A盒和B盒的结合通过直接相互作用在空间上阻碍了TFIIIB的锚定,导致TFIIIC - B - DNA复合物的稳定性降低。此外,这个低转录基因侧翼区域中存在的多个TATATAA序列成功竞争TFIIIB,从而减少转录。通过补充TFIIIB可以在一定程度上提高转录水平,但补充TATA盒结合蛋白则不能。因此,-6,7的低转录既归因于形成了不太稳定的转录复合物,也归因于TFIIIB的隔离。过量的转录因子TFIIIB的可用性可以作为一种普遍机制,根据组织内的发育需求启动基因家族所有个体成员的转录。