Tan S, Troxler R F
Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5316-21. doi: 10.1073/pnas.96.9.5316.
Two distinct cDNAs encoding putative sigma factors of plastid RNA polymerase were isolated from Zea mays, a C4 plant. The deduced amino acid sequences of both cDNAs possess all four highly conserved domains proposed for recognition of -10 and -35 promoter elements, core complex binding, DNA binding, and melting. These two cDNAs are designated sig1 and sig2. Phylogenetic analysis of available plastid sigma factors indicated that they were probably the descendants of cyanobacterial principal sigma factors. Southern blots probed with sig1 and sig2 revealed that both genes exist in the maize nuclear genome as single-copy genes, but low-stringency hybridization suggested the presence of a multigene family of maize plastid sigma factors. Transcription of sig1 and sig2 is light inducible and tissue specific. Transcripts of sig1 and sig2 were abundant in greening leaf tissues; sig2 (but not sig1) was barely detectable in etiolated leaves and neither was detectable in roots. Immunological studies using a peptide antibody against an epitope in subdomain 2.4 of Sig1 revealed 50-kDa and 60-kDa immunoreactive proteins in maize chloroplasts. Reduced levels of the 60-kDa immunoreactive protein were detected in etioplasts, and no immunoreactive proteins were observed in roots. Collectively, the data suggest that the nuclear genes, sig1 and sig2, may play a role in differential expression of plastid genes during chloroplast biogenesis.
从C4植物玉米中分离出了两个编码质体RNA聚合酶假定σ因子的不同cDNA。这两个cDNA推导的氨基酸序列均具有为识别-10和-35启动子元件、核心复合物结合、DNA结合和解链所提出的全部四个高度保守结构域。这两个cDNA分别命名为sig1和sig2。对现有质体σ因子的系统发育分析表明,它们可能是蓝藻主要σ因子的后代。用sig1和sig2进行Southern杂交显示,这两个基因在玉米核基因组中均以单拷贝基因形式存在,但低严谨度杂交表明存在玉米质体σ因子多基因家族。sig1和sig2的转录是光诱导的且具有组织特异性。sig1和sig2的转录本在叶片绿化组织中丰富;sig2(而非sig1)在黄化叶片中几乎检测不到,在根中均检测不到。使用针对Sig1 2.4亚结构域中一个表位的肽抗体进行的免疫学研究在玉米叶绿体中发现了50 kDa和60 kDa的免疫反应蛋白。在黄化质体中检测到60 kDa免疫反应蛋白水平降低,在根中未观察到免疫反应蛋白。总体而言,数据表明核基因sig1和sig2可能在叶绿体生物发生过程中质体基因的差异表达中起作用。