Abstract
Thermomonospora fusca cellulase E5 is encoded by the celE gene. This gene appears to be regulated at the transcriptional level by both induction and repression, and three putative closely linked promoters have been located by S1 mapping. To study its regulatory mechanism, a gel retardation assay was used to identify a protein in T. fusca cell extracts that interacted specifically with the DNA fragment containing the celE promoters. It was found that the binding activity appeared only when cellulase synthesis was induced, and it therefore resembled an activator protein involved in cellulase induction. DNase I footprinting identified the target sequence for this protein as a 21-base-pair sequence downstream from the putative celE promoters. The level of this protein was measured in two cellulase constitutive mutants, and the results suggest a complex control for celE induction.
MeSH Terms
Bacterial Proteins/analysis,genetics,metabolism
Base Sequence
Cellulase/biosynthesis,genetics
DNA, Bacterial/genetics,metabolism
DNA-Binding Proteins/analysis,genetics,metabolism
Deoxyribonuclease I
Gene Expression Regulation
Hot Temperature
Micromonosporaceae/enzymology,genetics
Molecular Sequence Data
Promoter Regions, Genetic
Chemicals
Bacterial Proteins
DNA, Bacterial
DNA-Binding Proteins
Deoxyribonuclease I
Cellulase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lin E S
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Wilson D B
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