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Multiple protein factors bind to a rice glutelin promoter region.


ABSTRACT: Our goal is to identify cis-acting elements in the regulatory region of the major seed storage protein gene in rice. A glutelin gene (pGL5-1) has been cloned by screening a rice genomic DNA library with synthetic oligonucleotides and with an amplified DNA fragment. A transient expression assay using immature rice seeds shows that its 5' flanking sequence can direct the synthesis of beta-glucuronidase (GUS) when fused upstream of the GUS coding region. Gel-retardation assays were performed to study protein-DNA interactions between putative regulatory sequences of pGL5-1 and nuclear proteins from immature rice seeds. We demonstrate that at least six protein-DNA complexes are formed between the 5' flanking sequence of pGL5-1 (-677 to -45) and nuclear protein factors. By subsequent DNase I-footprinting analyses we defined several protein-binding regions. Two of the protein-binding sequences contain the TGAGTCA motif, which is also present in the -300 element found in the 5' flanking sequences of several storage protein genes of other crop plants, and to which the transcription factors jun and GCN4 bind.

SUBMITTER: Kim SY 

PROVIDER: S-EPMC332740 | biostudies-other | 1990 Dec

REPOSITORIES: biostudies-other

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Multiple protein factors bind to a rice glutelin promoter region.

Kim S Y SY   Wu R R  

Nucleic acids research 19901201 23


Our goal is to identify cis-acting elements in the regulatory region of the major seed storage protein gene in rice. A glutelin gene (pGL5-1) has been cloned by screening a rice genomic DNA library with synthetic oligonucleotides and with an amplified DNA fragment. A transient expression assay using immature rice seeds shows that its 5' flanking sequence can direct the synthesis of beta-glucuronidase (GUS) when fused upstream of the GUS coding region. Gel-retardation assays were performed to stu  ...[more]

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