McCormac D J, Litz H, Wang J, Gollnick P D, Berry J O
Department of Biological Sciences, State University of New York, Buffalo, New York 14260, USA.
J Biol Chem. 2001 Feb 2;276(5):3476-83. doi: 10.1074/jbc.M009236200. Epub 2000 Nov 13.
In amaranth, a C(4) dicotyledonous plant, the plastid rbcL gene (encoding the large subunit of ribulose-1,5-bisphosphate carboxylase) is regulated post-transcriptionally during many developmental processes, including light-mediated development. To identify post-transcriptional regulators of rbcL expression, three types of analyses (polysome heel printing, gel retardation, and UV cross-linking) were utilized. These approaches revealed that multiple proteins interact with 5' regions of rbcL mRNA in light-grown, but not etiolated, amaranth plants. Light-associated binding of a 47-kDa protein (p47), observed by UV cross-linking, was highly specific for the rbcL 5' RNA. Binding of p47 occurred only with RNAs corresponding to mature processed rbcL transcripts (5'-untranslated region (UTR) terminating at -66); transcripts with longer 5'-UTRs did not associate with p47 in vitro. Variations in the length of the rbcL 5'-UTR were found to occur in vivo, and these different 5' termini may prevent or enhance light-associated p47 binding, possibly affecting rbcL expression as well. p47 binding correlates with light-dependent rbcL polysome association of the fully processed transcripts in photosynthetic leaves and cotyledons but not with cell-specific rbcL mRNA accumulation in bundle sheath and mesophyll chloroplasts.
在苋属植物(一种C4双子叶植物)中,质体rbcL基因(编码核酮糖-1,5-二磷酸羧化酶的大亚基)在包括光介导发育在内的许多发育过程中受到转录后调控。为了鉴定rbcL表达的转录后调节因子,采用了三种分析方法(多核糖体足迹分析、凝胶阻滞分析和紫外线交联分析)。这些方法表明,在光照生长而非黄化的苋属植物中,多种蛋白质与rbcL mRNA的5'区域相互作用。通过紫外线交联观察到的一种47 kDa蛋白质(p47)与光相关的结合,对rbcL 5' RNA具有高度特异性。p47的结合仅发生在与成熟加工的rbcL转录本相对应的RNA上(5'非翻译区(UTR)终止于-66);具有更长5'-UTR的转录本在体外不与p47结合。发现rbcL 5'-UTR的长度在体内存在变化,这些不同的5'末端可能会阻止或增强与光相关的p47结合,也可能影响rbcL的表达。p47的结合与光合叶片和子叶中完全加工的转录本的光依赖性rbcL多核糖体结合相关,但与维管束鞘和叶肉叶绿体中细胞特异性rbcL mRNA积累无关。