George Janet A, Pardue Mary-Lou
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Genetics. 2003 Feb;163(2):625-35. doi: 10.1093/genetics/163.2.625.
The Drosophila telomeric retrotransposon, HeT-A, is found only in heterochromatin; therefore, its promoter must function in this chromatin environment. Studies of position effect variegation suggest that promoters of heterochromatic genes are very different from euchromatic promoters, but this idea has not been tested with isolated promoter sequences. The HeT-A promoter is the first heterochromatin promoter to be isolated and it is of interest to investigate its activity when removed from telomeric heterochromatin. This promoter was initially characterized by testing reporter constructs in transient transfection of cultured cells, an environment that may approximate its endogenous heterochromatin. We now report P-element-mediated transpositions of these constructs, testing the function of different parts of the putative promoter in euchromatin. Expression of endogenous HeT-A RNA shows marked developmental regulation and accumulates preferentially in replicating diploid tissues. HeT-A promoter constructs are active in all euchromatic locations tested and some display aspects of endogenous HeT-A stage- and cell-type expression programs. The activity of each promoter construct in euchromatic locations is also generally consistent with its activity in the transient transfection tests; a possibly significant exception is one sequence segment that appreciably enhanced activity in transient transfection but repressed promoter activity in euchromatin.
果蝇端粒逆转座子HeT-A仅存在于异染色质中;因此,其启动子必须在这种染色质环境中发挥作用。位置效应斑驳的研究表明,异染色质基因的启动子与常染色质启动子非常不同,但这一观点尚未通过分离的启动子序列进行验证。HeT-A启动子是第一个被分离的异染色质启动子,研究其从端粒异染色质中移除后的活性很有意义。该启动子最初通过在培养细胞的瞬时转染中测试报告基因构建体来表征,这种环境可能接近其内源性异染色质。我们现在报告这些构建体的P元件介导的转座,测试推定启动子不同部分在常染色质中的功能。内源性HeT-A RNA的表达显示出明显的发育调控,并优先在复制的二倍体组织中积累。HeT-A启动子构建体在所有测试的常染色质位置均有活性,一些构建体显示出内源性HeT-A阶段和细胞类型表达程序的某些方面。每个启动子构建体在常染色质位置的活性通常也与其在瞬时转染测试中的活性一致;一个可能显著的例外是一个序列片段,它在瞬时转染中显著增强活性,但在常染色质中抑制启动子活性。