Lee Y R, Nagao R T, Key J L
Department of Botany, University of Georgia, Athens 30602.
Plant Cell. 1994 Dec;6(12):1889-97. doi: 10.1105/tpc.6.12.1889.
A cDNA clone encoding a 101-kD heat shock protein (HSP101) of soybean was isolated and sequenced. Genomic DNA gel blot analysis indicated that the corresponding gene is a member of a multigene family. The mRNA for HSP101 was not detected in 2-day-old etiolated soybean seedlings grown at 28 degrees C but was induced by elevated temperatures. DNA sequence comparison has shown that the corresponding gene belongs to the Clp (caseinolytic protease) (or Hsp100) gene family, which is evolutionarily conserved and found in both prokaryotes and eukaryotes. On the basis of the spacer length between the two conserved ATP binding regions, this gene has been identified as a member of the ClpB subfamily. Unlike other Clp genes previously isolated from higher plants, the expression of this soybean Hsp101 gene is heat inducible, and it does not have an N-terminal signal peptide for targeting to chloroplasts. Transformation of the soybean Hsp101 gene into a yeast HSP104 deletion mutant complemented restoration of acquired thermotolerance, a process in which cells survive an otherwise lethal heat stress after they are given a permissive heat treatment.
分离并测序了一个编码大豆101-kD热休克蛋白(HSP101)的cDNA克隆。基因组DNA凝胶印迹分析表明,相应的基因是一个多基因家族的成员。在28℃下生长的2日龄黄化大豆幼苗中未检测到HSP101的mRNA,但高温可诱导其产生。DNA序列比较表明,相应的基因属于Clp(酪蛋白水解蛋白酶)(或Hsp100)基因家族,该家族在进化上保守,存在于原核生物和真核生物中。根据两个保守的ATP结合区域之间的间隔长度,该基因被鉴定为ClpB亚家族的成员。与先前从高等植物中分离的其他Clp基因不同,该大豆Hsp101基因的表达是热诱导型的,并且它没有用于靶向叶绿体的N端信号肽。将大豆Hsp101基因转化到酵母HSP104缺失突变体中,可补充获得性耐热性的恢复,在这个过程中,细胞在经过允许的热处理后能够在原本致命的热应激中存活下来。