Drzymalla C, Schroda M, Beck C F
Institut für Biologie III, Universität Freiburg, Germany.
Plant Mol Biol. 1996 Sep;31(6):1185-94. doi: 10.1007/BF00040835.
The nuclear heat shock gene HSP70B of Chlamydomonas reinhardtii is inducible by heat stress and light. Induction by either environmental cue resulted in a transient elevation in HSP70B protein. Here we describe the organization and nucleotide sequence of the HSP70B gene. The deduced protein exhibits a distinctly higher homology to prokaryotic HSP70s than to those of eukaryotes, including the cytosolic HSP70A of Chlamydomonas reinhardtii. The HSP70B protein, as previously demonstrated by in vitro translation, is synthesized with a cleavable presequence. Using an HSP70B-specific antibody, this heat shock protein was localized to the chloroplast by cell fractionation experiments. A stromal location was suggested by the presence of a conserved sequence motif used for cleavage of presequences by a signal peptidase of the stroma. Amino acid alignments of HSP70 proteins from various organisms and different cellular compartments allowed the identification of sequence motifs, which are diagnostic for HSP70s of chloroplasts and cyanobacteria.
莱茵衣藻的核热休克基因HSP70B可被热应激和光照诱导。这两种环境信号的诱导都会导致HSP70B蛋白短暂升高。在此,我们描述了HSP70B基因的结构和核苷酸序列。推导的蛋白与原核生物HSP70的同源性明显高于与真核生物HSP70的同源性,包括莱茵衣藻的胞质HSP70A。如先前体外翻译所证明的,HSP70B蛋白是由一个可切割的前导序列合成的。通过细胞分级分离实验,使用HSP70B特异性抗体将这种热休克蛋白定位到叶绿体中。基质中存在一个用于被基质信号肽酶切割前导序列的保守序列基序,这表明该蛋白位于基质中。对来自各种生物体和不同细胞区室的HSP70蛋白进行氨基酸比对,从而鉴定出了可用于诊断叶绿体和蓝细菌HSP70的序列基序。