Global analysis of transcription kinetics during competence development in Streptococcus pneumoniae using high density DNA arrays.
The kinetics of global changes in transcription patterns during competence development in Streptococcus pneumoniae was analysed with high-density arrays. Four thousand three hundred and one clones of a S. pneumoniae library, covering almost the entire genome, were amplified by PCR and gridded at high density onto nylon membranes. Competence was induced by the addition of CSP (competence stimulating peptide) to S. pneumoniae cultures grown to the early exponential phase. RNA was extracted from samples at 5 min intervals (for a period of 30 min) after the addition of CSP. Radiolabelled cDNA was generated from isolated total RNA by random priming and the probes were hybridized to identical high density arrays. Genes whose transcription was induced or repressed during competence were identified. Most of the genes previously known to be competence induced were detected together with several novel genes that all displayed the characteristic transient kinetics of competence-induced genes. Among the newly identified genes many have suggested functions compatible with roles in genetic transformation. Some of them may represent new members of the early or late competence regulons showing competence specific consensus sequences in their promoter regions. Northern experiments and mutational analysis were used to confirm some of the results.
Department of Microbiology, Glaxo Wellcome S.p.A., Verona, Italy.
R
R
Rimini
B
B
Jansson
G
G
Feger
T C
TC
Roberts
M
M
de Francesco
A
A
Gozzi
F
F
Faggioni
E
E
Domenici
D M
DM
Wallace
N
N
Frandsen
A
A
Polissi
D001483:Base Sequence; D015152:Blotting, Northern; D004269:DNA, Bacterial; D015536:Down-Regulation; D020869:Gene Expression Profiling / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000706:statistics & numerical data; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic; D015854:Up-Regulation
D004269:DNA, Bacterial