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In vitro transcription of PrfA‐dependent and
‐independent genes of Listeria monocytogenes
Author(s) -
LalicMülthaler M.,
Bohne J.,
Goebel W.
Publication year - 2001
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2001.02607.x
Subject(s) - promoter , biology , rna polymerase ii , rna polymerase , transcription (linguistics) , sigma factor , transcription factor ii d , microbiology and biotechnology , transcription factor ii b , gene , general transcription factor , transcription factor , gene expression , rna , genetics , linguistics , philosophy
In vitro transcription starting from the promoters of the Listeria monocytogenes genes hly , plcA , actA , mpl , prfA and iap has been studied. Whereas transcription from P hly , P plcA and P actA is strictly PrfA‐dependent, that from P iap , P prfA1/2 and, unexpectedly, also from P mpl is independent. Initiation of in vitro transcription at all tested promoters except P prfA requires high concentrations of ATP but not GTP. The nucleotides required in higher concentrations for efficient in vitro transcription are always included in the first three nucleotides of the corresponding transcript . RNA polymerase prepared from L. monocytogenes cultured either in rich culture medium (RNAP BHI ), exposed to heat shock conditions (RNAP 48 ) or conditioned in minimal essential medium (RNAP MEM ) shows significant differences in the transcription efficiencies when transcription is initiated at these promoters. Transcription starting from the PrfA‐dependent promoters P actA and P hly is enhanced with RNAP 48 and RNAP MEM (in relation to P iap –mediated transcription), while transcription from the other promoters is reduced when compared with RNAP BHI . These data suggest that in vivo transcription of the genes actA and hly may not function optimally with RNA polymerase loaded with the vegetative sigma factor 43, but may require a modified RNA polymerase, possibly loaded with an alternative sigma factor.