Regulation of competence for genetic transformation in Streptococcus pneumoniae: expression of dpnA, a late competence gene encoding a DNA methyltransferase of the DpnII restriction system

2000 ◽  
Vol 35 (5) ◽  
pp. 1089-1098 ◽  
Author(s):  
Sanford A. Lacks ◽  
Sahlu Ayalew ◽  
Adela G. de la Campa ◽  
Bill Greenberg
2009 ◽  
Vol 191 (10) ◽  
pp. 3359-3366 ◽  
Author(s):  
Andrew Piotrowski ◽  
Ping Luo ◽  
Donald A. Morrison

ABSTRACT Competence for genetic transformation in Streptococcus pneumoniae is a transient physiological state whose development is coordinated by a peptide pheromone (CSP) and its receptor, which activates transcription of two downstream genes, comX and comW, and 15 other “early” genes. ComX, a transient alternative sigma factor, drives transcription of “late” genes, many of which are essential for transformation. In vivo, ComW both stabilizes ComX against proteolysis by the ClpE-ClpP protease and stimulates its activity. Interestingly, stabilization of ComX by deletion of the gene encoding the ClpP protease did not extend the period of competence. We considered the hypothesis that the rapid decay of competence arises from a rapid loss of ComW and thus of its ComX stimulating activity, so that ComX might persist but lose its transcriptional activity. Western analysis revealed that ComW is indeed a transient protein, which is also stabilized by deletion of the gene encoding the ClpP protease. However, stabilizing both ComX and ComW did not prolong either ComX activity or the period of transformation, indicating that termination of the transcriptional activity of ComX is not dependent on proteolysis of ComW.


1991 ◽  
Vol 37 (9) ◽  
pp. 713-715 ◽  
Author(s):  
Vijay M. Chauthaiwale ◽  
Pranav R. Vyas ◽  
Vasanti V. Deshpande

A PEG-mediated transformation system for Chainia (NCL 82-5-1) was develolped using a broad host range Streptomyces vector, pIJ702. Protoplasts prepared from Chainia (NCL 82-5-1) were regenerated with 5% efficiency. Transformation of the protoplasts with pIJ702 gave 10–20 transformants/μg DNA. The low efficiency of transformation is attributed to a restriction system in Chainia; this could be inhibited by treating the protoplasts at 42 °C for 10 min just before transformation. The yield of transformants increased 100-fold when pIJ702 was modified by passage in Chainia. Because the plasmid replicon was functional in Chainia and the modified plasmid was stably maintained, the transformation system should be useful for self-cloning in Chainia NCL 82-5-1 of the many commercially important enzymes this strain is known to produce. Key words: Chainia, transformation, Streptomyces, pIJ702 restriction modification, heat attenuation.


Gene ◽  
1995 ◽  
Vol 157 (1-2) ◽  
pp. 209-212 ◽  
Author(s):  
Sanford A. Lacks ◽  
Bill Greenberg ◽  
Alexander G. Sabelnikov

PLoS Genetics ◽  
2013 ◽  
Vol 9 (9) ◽  
pp. e1003819 ◽  
Author(s):  
Calum Johnston ◽  
Stéphanie Caymaris ◽  
Aldert Zomer ◽  
Hester J. Bootsma ◽  
Marc Prudhomme ◽  
...  

Gene ◽  
1997 ◽  
Vol 194 (2) ◽  
pp. 249-255 ◽  
Author(s):  
Yian-Biao Zhang ◽  
Bill Greenberg ◽  
Sanford A Lacks

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