Residual polyadenylation in poly(A) polymerase I (pcnB ) mutants of Escherichia coli does not result from the activity encoded by the f310 gene
Open Access
- 1 December 1999
- journal article
- research article
- Published by Wiley in Molecular Microbiology
- Vol. 34 (5) , 1109-1119
- https://doi.org/10.1046/j.1365-2958.1999.01674.x
Abstract
As extracts of poly(A) polymerase I (PAP I) deficient strains of Escherichia coli appeared to contain considerable residual polyadenylating activity, efforts were undertaken to identify a second poly(A) polymerase. Recently, a gene (f310 ) encoding the putative second poly(A) polymerase was cloned and sequenced. Here we have tested the ability of the F310 protein to add poly(A) tails in vivo by measuring total poly(A) levels in both f310 mutants and strains that overproduce F310. In addition, we have visualized poly(A) tails and examined ColE1 plasmid copy number in various genetic backgrounds. We also carried out direct biochemical measurements of AMP incorporation, using cell extracts after amplification of F310. All the data obtained indicate that F310 is not a poly(A) polymerase. Although the presence of two potential ATP binding domains in the F310 protein may account for its apparent ATP binding activity, its true biochemical function remains to be identified. In addition, we show that the f310 gene is transcribed, almost exclusively, during stationary phase from a σs promoter.Keywords
This publication has 25 references indexed in Scilit:
- Differential Sensitivities of Portions of the mRNA for Ribosomal Protein S20 to 3′-Exonucleases Dependent on Oligoadenylation and RNA Secondary StructureJournal of Biological Chemistry, 1996
- Polyadenylylation helps regulate mRNA decay in Escherichia coli.Proceedings of the National Academy of Sciences, 1995
- Identification of a Second Poly(A) Polymerase in Escherichia coliBiochemical and Biophysical Research Communications, 1994
- PcnB is required for the rapid degradation of RNAI, the antisense RNA that controls the copy number of CoIE1‐related plasmidsMolecular Microbiology, 1993
- The Escherichia coli pcnB gene promotes adenylylation of antisense RNAI of ColE1-type plasmids in vivo and degradation of RNAI decay intermediates.Proceedings of the National Academy of Sciences, 1993
- Baculovirus vectors for multiple gene expression and for occluded virus productionGene, 1991
- Analysis of nutR, a site required for transcription antitermination in phage lambda.Proceedings of the National Academy of Sciences, 1987
- Mutations in a new chromosomal gene of Escherichia coli K-12, pcnB, reduce plasmid copy number of pBR322 and its derivativesMolecular Genetics and Genomics, 1986
- 17 Polynucleotide PhosphorylasePublished by Elsevier ,1982
- Complete Nucleotide Sequence of the Escherichia coli Plasmid pBR322Published by Cold Spring Harbor Laboratory ,1979