Inhibitory effect of adenosine 3',5'-phosphate on cell division of Escherichia coli K-12 mutant derivatives

Abstract

Cell division of Escherichia coli K-12 strain PA3092 was inhibited by the addition of adenosine 3',5'-phosphate (cAMP), and the cellular morphology was changed from rods into flaments. Nucleoids in the filaments were regularly distributed and septum formation was perfectly inhibited. This inhibition of cell division by cAMP was reversed by the addition of guanosine 3',5'-monophosphate. To examine whether the inhibitory effect of cAMP on cell division in E. coli PA3092 was specific, its effect in several parental strains was investigated. Induction of cell filamentation by cAMP was observed in E. coli PA309 and P678, but not in E. coli W595, Wl, Y10, or the wild-type strain. This result suggests that filamentation by cAMP in E. coli PA3092, PA309, and P678 was due to the mutagenesis by which E. coli P678 was derived from E. coli W595.Cyclic AMP (cAMP) exerts its effect in Escherichia coli primarily by stimulating the synthesis of a number of proteins (19). The most important example is the control of the synthesis of ,B-galactosidase, the gene of which is transcribed at a significant rate in the presence of cAMP and cAMP receptor protein (CRP). In addition to controlling the synthesis of these proteins positively (positive control), cAMP has been found to regulate the synthesis of glutamate synthase, glutaminase A (18), and the outer membrane protein III (15) negatively (negative control).On the other hand, it has been reported that adenylate cyclase-deficient (cya) mutants and CRP-deficient (crp) mutants show cocci or short rod morphology (11,13), and the increase in intracellular cAMP concentration precedes the observed change in cellular morphology from cocci to rods in Arthrobacter crystallopoites (10). Furthermore, Aono et al. reported that synthesis of several envelope proteins in E. coli is regulated by cAMP and CRP (2). These results suggest that there is some correlation between cAMP and cell morphology.Recently, we found that in E. coli PA3092, rods were changed into filaments upon the addition of cAMP. There have been many reports about filamentation in E. coli (7-9, 12, 23-26), but filamentation by cAMP has not been reported yet. It is important if cell division is controlled by cAMP, which is a regulatory molecule in transcription. Several temperature-sen-sitive cell division mutants have been isolated (17) and used to investigate cell division in E. coli, but the control mechanism of cell division is poorly understood. Studies with E. coli PA3092 should provide useful information regarding the cell division process in E. coli. This report presents characteristics of the inhibition of cell division and the morphological change in E. coli caused by cAMP and suggests the control mechanism of filamentation involving this molecule.

Identification and characterization of the Escherichia coli envC gene encoding a periplasmic coiled-coil protein with putative peptidase activity

“…A transduction experiment using a cysE: :Tn5 derivative of MG1655 (JE7989) as a recipient gave essentially the same result ( Fig. 1E^G), indicating the irrelevance of the ¢c (¢lamentation induced by cAMP) mutation at min 75.2 in P678 [25,26]. All the envC transductants were inhibited by 10 Wg ml 31 crystal violet.…”

Involvement of cyclic AMP and its receptor protein in filamentation of an Escherichia coli fic mutant

“…Protein concentration was determined by the method of Lowry et al with bovine serum albumin as the standard (14). the presence of 10 mM cAMP (20). The inhibitory effect of cAMP was not observed in a crp mutant, E. coli 5333 (data not shown).…”

“…Recently, we found that E. coli PA3092 grew into filaments on the addition of cyclic AMP (cAMP) at an elevated temperature (43°C) and that the filamentation was reversed by cyclic GMP, which is known to be a competitive ligand for the cAMP receptor protein (CRP) (20). The evidence suggests that cell division could be controlled by a cAMP-CRP complex.…”

“…Media and buffer. E. coli was grown in L broth medium (20). Eosin methylene blue (EMB) plates were used to investigate the fermentation of rhamnose and glycerol.…”

“…L broth and EMB plates were solidified with 1.5% agar. The starvation buffer has been described (20). Medium A and Z buffers were used for assay of 1galactosidase (15).…”