Kim Yu Jung, Hice Robert H, O'Brochta David A, Atkinson Peter W
Graduate Program in Department of Biochemistry and Molecular Biology, University of California, Riverside, CA 92521, USA.
Genetica. 2011 Aug;139(8):985-97. doi: 10.1007/s10709-011-9600-2. Epub 2011 Jul 31.
We have conducted a structure and functional analysis of the hobo transposable element of Drosophila melanogaster. A minimum of 141 bp of the left (L) end and 65 bp of the right (R) end of the hobo were shown to contain sequences sufficient for transposition. Both ends of hobo contain multiple copies of the motifs GGGTG and GTGGC and we show that the frequency of hobo transposition increases as a function of the copy number of these motifs. The R end of hobo contains a unique 12 bp internal inverted repeat that is identical to the hobo terminal inverted repeats. We show that this internal inverted repeat suppresses transposition activity in a hobo element containing an intact L end and only 475 bp of the R end. In addition to establishing cis-sequences requirements for transposition, we analyzed trans-sequence effects of the hobo transposase. We show a hobo transposase lacking the first 49 amino acids catalyzed hobo transposition at a higher frequency than the full-length transposase suggesting that, similar to the related Ac transposase, residues at the amino end of the transposase reduce transposition. Finally, we compared target site sequences of hobo with those of the related Hermes element and found both transposons have strong preferences for the same insertion sites.
我们对黑腹果蝇的hobo转座元件进行了结构和功能分析。结果显示,hobo元件左端(L端)至少141 bp和右端(R端)65 bp的序列包含足以进行转座的序列。hobo元件的两端都含有多个GGGTG和GTGGC基序拷贝,并且我们发现hobo转座频率随着这些基序拷贝数的增加而升高。hobo元件的R端含有一个独特的12 bp内部反向重复序列,该序列与hobo元件的末端反向重复序列相同。我们发现,这个内部反向重复序列会抑制一个含有完整L端和仅475 bp R端的hobo元件的转座活性。除了确定转座所需的顺式序列外,我们还分析了hobo转座酶的反式序列效应。我们发现,缺少前49个氨基酸的hobo转座酶催化hobo转座的频率高于全长转座酶,这表明,与相关的Ac转座酶类似,转座酶氨基端的残基会降低转座频率。最后,我们将hobo元件的靶位点序列与相关的Hermes元件的靶位点序列进行了比较,发现这两种转座子对相同的插入位点都有很强的偏好。