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J. Biol. Chem., Vol. 276, Issue 5, 3476-3483, February 2, 2001
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,
From the Department of Biological Sciences, State University of New
York, Buffalo, New York 14260
In amaranth, a C4
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.
Present address: Imaging Research, Inc., Brock University, St.
Catherines, ON, CA.
§
To whom correspondence should be addressed. Tel.: 716-645-3488;
Fax: 716-645-2975; E-mail: camjob@acsu.buffalo.edu.
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