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Phosphodiesterase expression and cyclic di‐GMP production in Streptomyces coelicolor (1013.22)
Author(s) -
Geiger Robert,
Tansey John,
Bennett Jennifer
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1013.22
Subject(s) - streptomyces coelicolor , phosphodiesterase , gene , gene expression , mutant , fusion gene , biology , microbiology and biotechnology , promoter , streptomyces , chemistry , bacteria , biochemistry , genetics , enzyme
Streptomyces coelicolor is a pharmacologically important gram‐positive, filamentous, soil bacterium that displays a complex pattern of differentiation. The second messenger cyclic di‐GMP has been measured through the bacterium’s life‐cycle. We hypothesize that phosphodiesterases, which break down cyclic di‐GMP are differentially expressed to control developmental progression. We have constructed mutants that have a transposon insertion containing the gene for EGFP (enhanced green fluoresent protein) that disrupts a phosphodiesterase gene and forms a transcriptional fusion to the native phosphodiesterase promotor. EGFP was directly measured via ELISA and quantitated using fluorescence. Expression from the promotor for two phosphodiesterase genes varies between 36 and 96 hours. A plunge in EGFP expression is observed before the seventy‐two hour growth point which correlates with maximum sporulation in the wild type strain. In addition, cyclic di‐GMP production has been measured during the bacterium’s life cycle using HPLC‐MS/MS. The data shows differential production of the di nucleotide. These data provide further evidence that cyclic di‐GMP plays a role in the developmental progression of S. coelicolor.