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J Biol Chem ; 276(5): 3476-83, 2001 Feb 02.
Article in English | MEDLINE | ID: mdl-11076953

ABSTRACT

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.


Subject(s)
5' Untranslated Regions/metabolism , Blood Proteins/physiology , Chloroplasts/metabolism , Magnoliopsida/metabolism , Phosphoproteins/physiology , Plant Proteins/metabolism , Ribulose-Bisphosphate Carboxylase , 5' Untranslated Regions/chemistry , 5' Untranslated Regions/radiation effects , Blood Proteins/metabolism , Chloroplasts/genetics , Gene Expression Regulation, Plant/radiation effects , Light , Magnoliopsida/genetics , Phosphoproteins/metabolism , Plant Proteins/genetics , Plant Proteins/radiation effects , Polyribosomes/metabolism , RNA, Messenger/metabolism , RNA, Messenger/radiation effects , RNA-Binding Proteins/isolation & purification , RNA-Binding Proteins/metabolism , Ultraviolet Rays
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