Lohe A R, Sullivan D T, Hartl D L
Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Genetics. 1996 Nov;144(3):1087-95. doi: 10.1093/genetics/144.3.1087.
We have studied the Mos1 transposase encoded by the transposable element mariner. This-transposase is a member of the "D,D(35)E" superfamily of proteins exhibiting the motif D,D(34)D. It is not known whether this transposase, or other eukaryote transposases manifesting the D,D(35)E domain, functions in a multimeric form. Evidence for oligomerization was found in the negative complementation of Mos1 by an EMS-induced transposase mutation in the catalytic domain. The transposase produced by this mutation has a glycine-to-arginine replacement at position 292. The G292R mutation strongly interferes with the ability of wild-type transposase to catalyze excision of a target element. Negative complementation was also observed for two other EMS mutations, although the effect was weaker than observed with G292R. Results from the yeast two-hybrid system also imply that Mos1 subunits interact, suggesting the possibility of subunit oligomerization in the transposition reaction. Overproduction of Mos1 subunits through an hsp70 promoter also inhibits excision of the target element, possibly through autoregulatory feedback on transcription or through formation of inactive or less active oligomers. The effects of both negative complementation and overproduction may contribute to the regulation of mariner transposition.
我们研究了转座元件水手座编码的Mos1转座酶。这种转座酶是表现出D,D(34)D基序的“D,D(35)E”蛋白质超家族的成员。目前尚不清楚这种转座酶,或其他表现出D,D(35)E结构域的真核转座酶是否以多聚体形式发挥作用。在催化结构域中由EMS诱导的转座酶突变对Mos1的负互补中发现了寡聚化的证据。这种突变产生的转座酶在第292位有一个甘氨酸到精氨酸的替换。G292R突变强烈干扰野生型转座酶催化靶元件切除的能力。另外两个EMS突变也观察到了负互补,尽管其效果比G292R弱。酵母双杂交系统的结果也表明Mos1亚基相互作用,这表明在转座反应中存在亚基寡聚化的可能性。通过hsp70启动子过量表达Mos1亚基也会抑制靶元件的切除,可能是通过对转录的自调节反馈或通过形成无活性或活性较低的寡聚体。负互补和过量表达的影响可能都有助于调节水手座转座。