Suoniemi A, Narvanto A, Schulman A H
Institute of Biotechnology, University of Helsinki, Finland.
Plant Mol Biol. 1996 May;31(2):295-306. doi: 10.1007/BF00021791.
The BARE-1 retrotransposon occurs in more than 10(4) copies in the barley genome. The element is bounded by long terminal repeats (LTRs, 1829 bp) containing motifs typical of retrotransposon promoters. These, the presence of predicted priming sites for reverse transcription, and the high conservation for all key functional domains of the coding region suggest that copies within the genome could be active retrotransposons. In view of this, we looked for transcription of BARE-1 within barley tissues and examined the promoter function of the BARE-1 LTR. We demonstrate here that BARE-1-like elements are transcribed in barley tissues, and that the transcripts begin within the BARE-1 LTR downstream of TATA boxes. The LTR can drive expression of reporter genes in transiently transformed barley protoplasts. This is dependent on the presence of a TATA box functional in planta as well. Furthermore, we identify regions within the LTR responsible for expression within protoplasts by deletion analyses of LTR-luc constructs. Similarities between promoter regulatory motifs and regions of the LTR were identified by comparisons to sequence libraries. The activity of the LTR as a promoter, combined with the abundance of BARE-1 in the genome, suggests that BARE-1 may retain the potential for propagation in the barley genome.
BARE-1反转录转座子在大麦基因组中的拷贝数超过10⁴个。该元件由长末端重复序列(LTRs,1829 bp)界定,这些长末端重复序列包含反转录转座子启动子的典型基序。这些基序、预测的反转录起始位点的存在以及编码区所有关键功能域的高度保守性表明,基因组中的拷贝可能是活跃的反转录转座子。鉴于此,我们在大麦组织中寻找BARE-1的转录情况,并检测了BARE-1 LTR的启动子功能。我们在此证明,大麦组织中存在类似BARE-1的元件转录,且转录本起始于TATA框下游的BARE-1 LTR内。该LTR可在瞬时转化的大麦原生质体中驱动报告基因的表达。这同样依赖于植物中起作用的TATA框的存在。此外,我们通过对LTR-luc构建体的缺失分析,确定了LTR内负责原生质体中表达的区域。通过与序列文库比较,鉴定出了启动子调控基序与LTR区域之间的相似性。LTR作为启动子的活性,加上基因组中BARE-1的丰富性,表明BARE-1可能在大麦基因组中保留了增殖的潜力。