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Identification of multicopy suppressors of the pcnB plasmid copy number defect in Escherichia coli.
Mol Genet Genomics 2002; 268(1):62-9MG

Abstract

Plasmids containing a ColE1 origin of replication are widely used for cloning purposes in Escherichia coli. Among the host factors that affect the copy number of ColE1 plasmids is the E. coli protein poly(A) polymerase I (PAP I), which regulates the intracellular level of RNA I, a ColE1-encoded negative regulator of plasmid replication. In strains that lack PAP I, RNA I levels are elevated, resulting in reduced levels of ColE1 plasmids in the cell. PAP I is encoded by the gene pcnB. We devised a genetic approach, based on the identification of multicopy suppressor clones, to identify trans-acting factors that can help offset the ColE1 plasmid copy number defect in a pcnB (-) genetic background. Using this strategy, we identified suppressors that mapped to two regions of the E. coli chromosome. The suppressor activity of one of the chromosomal regions was localized to the rssB gene, a response regulator gene known to be involved in the turnover of the stationary-phase sigma factor, RpoS. The second suppressor maps to min 55.4 of the E. coli chromosome, and the factor responsible for the suppressor activity appears to be a novel RNA or protein.

Authors+Show Affiliations

Boston Biomedical Research Institute, 64 Grove Street, Watertown, MA 02472, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

12242500

Citation

Sarkar, N, et al. "Identification of Multicopy Suppressors of the pcnB Plasmid Copy Number Defect in Escherichia Coli." Molecular Genetics and Genomics : MGG, vol. 268, no. 1, 2002, pp. 62-9.
Sarkar N, Cao GJ, Jain C. Identification of multicopy suppressors of the pcnB plasmid copy number defect in Escherichia coli. Mol Genet Genomics. 2002;268(1):62-9.
Sarkar, N., Cao, G. J., & Jain, C. (2002). Identification of multicopy suppressors of the pcnB plasmid copy number defect in Escherichia coli. Molecular Genetics and Genomics : MGG, 268(1), pp. 62-9.
Sarkar N, Cao GJ, Jain C. Identification of Multicopy Suppressors of the pcnB Plasmid Copy Number Defect in Escherichia Coli. Mol Genet Genomics. 2002;268(1):62-9. PubMed PMID: 12242500.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of multicopy suppressors of the pcnB plasmid copy number defect in Escherichia coli. AU - Sarkar,N, AU - Cao,G-J, AU - Jain,C, Y1 - 2002/08/13/ PY - 2002/01/04/received PY - 2002/06/20/accepted PY - 2002/9/21/pubmed PY - 2002/11/26/medline PY - 2002/9/21/entrez SP - 62 EP - 9 JF - Molecular genetics and genomics : MGG JO - Mol. Genet. Genomics VL - 268 IS - 1 N2 - Plasmids containing a ColE1 origin of replication are widely used for cloning purposes in Escherichia coli. Among the host factors that affect the copy number of ColE1 plasmids is the E. coli protein poly(A) polymerase I (PAP I), which regulates the intracellular level of RNA I, a ColE1-encoded negative regulator of plasmid replication. In strains that lack PAP I, RNA I levels are elevated, resulting in reduced levels of ColE1 plasmids in the cell. PAP I is encoded by the gene pcnB. We devised a genetic approach, based on the identification of multicopy suppressor clones, to identify trans-acting factors that can help offset the ColE1 plasmid copy number defect in a pcnB (-) genetic background. Using this strategy, we identified suppressors that mapped to two regions of the E. coli chromosome. The suppressor activity of one of the chromosomal regions was localized to the rssB gene, a response regulator gene known to be involved in the turnover of the stationary-phase sigma factor, RpoS. The second suppressor maps to min 55.4 of the E. coli chromosome, and the factor responsible for the suppressor activity appears to be a novel RNA or protein. SN - 1617-4615 UR - https://www.unboundmedicine.com/medline/citation/12242500/Identification_of_multicopy_suppressors_of_the_pcnB_plasmid_copy_number_defect_in_Escherichia_coli_ L2 - https://dx.doi.org/10.1007/s00438-002-0723-0 DB - PRIME DP - Unbound Medicine ER -