Abstract/Details

Isolation and characterization of proteins associated with the maize G-box binding complex recognizing a promoter element of the Adh1 gene

De Vetten, Nicolaas Clemens M. H.   University of Florida ProQuest Dissertations Publishing,  1993. 9505751.

Abstract (summary)

The G-box element is a moderately conserved component of the promoter of many inducible genes including the alcohol dehydrogenase genes of Arabidopsis and maize. To characterize maize proteins that are part of the G-box binding complex, monoclonal antibodies were generated against partially purified G-box binding factor (GBF) activity. Antibodies interact with partially purified maize GBF complexes to produce a slower migrating complex in the electrophoretic mobility shift assay (EMSA). Immunoprecipitation experiments suggest that the protein recognized by the antibody is not a DNA binding protein in and of itself, but rather is associated with a 46 kDa GBF. These monoclonal antibodies were used to isolate two classes of cDNA clones encoding proteins that we have designated GF14. The deduced amino acid sequence of maize GF14 is more than 60% identical to a class of mammalian brain proteins described as mediators of protein kinase C activity.

The sequence of the genes encoding the two GF14 mRNAs has been determined and contains five introns. The transcription units of both genes were found to be very similar with complete conservation of the intron position and length. The $5\sp\prime$ upstream region of both GF14 genes revealed the presence of a conserved G-box element. EMSA of maize GBF protein extract with the GF14 G-box indicates that GBF is binding to this site in the $5\sp\prime$ upstream region of GF14. GF14 protein is associated with the G-box binding complex interacting with the GF14 upstream region, suggesting that GF14 gene transcription is autoregulated.

To further characterize the maize G-box binding complex, a cDNA encoding maize GBF was isolated. The deduced amino acid sequence revealed conserved basic and leucine zipper domains characteristic of transcription factors of the bZIP class. GBF mRNA accumulates rapidly in cells exposed to anaerobic conditions, and the timing of accumulation precedes that of Adh1 mRNA accumulation. Furthermore, GBF binds to the minimal anaerobic-responsive promoter region of the Adh1 gene. This suggests that GBF is involved in anaerobic upregulation of Adh1 gene expression.

Indexing (details)


Subject
Molecular biology
Classification
0307: Molecular biology
Identifier / keyword
Biological sciences
Title
Isolation and characterization of proteins associated with the maize G-box binding complex recognizing a promoter element of the Adh1 gene
Author
De Vetten, Nicolaas Clemens M. H.
Number of pages
150
Degree date
1993
School code
0070
Source
DAI-B 55/09, Dissertation Abstracts International
Place of publication
Ann Arbor
Country of publication
United States
ISBN
979-8-208-03855-0
Advisor
Ferl, Robert J.
University/institution
University of Florida
University location
United States -- Florida
Degree
Ph.D.
Source type
Dissertation or Thesis
Language
English
Document type
Dissertation/Thesis
Dissertation/thesis number
9505751
ProQuest document ID
304063301
Copyright
Database copyright ProQuest LLC; ProQuest does not claim copyright in the individual underlying works.
Document URL
https://www.proquest.com/docview/304063301