AT-rich promoter elements of soybean heat shock gene Gmhsp17.5E bind two distinct sets of nuclear proteins in vitro.
A 33 bp double-stranded oligonucleotide homologous to two AT-rich sequences located upstream (-907 to -889 and -843 to -826) to the start of transcription of heat shock gene Gmhsp17.5E of soybean stimulated transcription when placed 5' to a truncated (-140) maize Adh1 promoter. The chimeric promoter was assayed in vivo utilizing anaerobically stressed sunflower tumors transformed by a pTi-based vector of Agrobacterium tumefaciens. Nuclear proteins extracted from soybean plumules were shown to bind double-stranded oligonucleotides homologous to AT-rich sequences in the 5' flanking regions of soybean beta-conglycinin, lectin, leghemoglobin and heat shock genes. These proteins were also shown to bind AT-rich probes homologous to homeobox protein binding sites from the Antennapedia and engrailed/fushi tarazu genes of Drosophila. Binding activity specific for AT-rich sequences showed a wide distribution among various plant organs and species. Preliminary characterization indicated that two sets of nuclear proteins from soybean bind AT-rich DNA sequences: a diverse high-molecular-weight (ca. 46-69 kDa) group, and a low-molecular-weight (23 and 32 kDa) group of proteins. The latter meets the operational criteria for high-mobility group proteins (HMGs).
Czarnecka
E
E
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611-0100.
Ingersoll
J C
JC
Gurley
W B
WB
D000426:Alcohol Dehydrogenase / Q000235:genetics; D001482:Base Composition; D001483:Base Sequence; D003001:Cloning, Molecular; D006360:Heat-Shock Proteins / Q000235:genetics*; D006609:High Mobility Group Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013025:Glycine max / Q000235:genetics*; D003313:Zea mays / Q000201:enzymology / Q000235:genetics
D006360:Heat-Shock Proteins; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; D000426:Alcohol Dehydrogenase