Abstract
F-like plasmids require a number of genes for conjugation, including tra operon genes and genes traM and traJ, which lie outside the tra operon. We now establish that a gene in the "leading region," gene 19, provides an important function during conjugation and RNA phage infection. Mutational inactivation of gene 19 on plasmid R1-16 by introduction of two nonpolar stop codons results in a 10-fold decrease in the conjugation frequency. Furthermore, infection studies with the male-specific bacteriophage R17 revealed that the phage is not able to form clear plaques in Escherichia coli cells carrying an R1-16 plasmid with the defective copy of gene 19. The total number of cells infected by phage R17 is reduced by a factor of 10. Both the conjugation- and infection-attenuated phenotypes caused by the defective gene 19 can be complemented in trans by introducing gene 19 alleles encoding the wild-type protein. Restoration of the normal phenotypes is also possible by introduction of the pilT gene encoded by the unrelated IncI plasmid R64. Our functional studies and similarities of protein 19 to proteins encoded by other DNA transfer systems, as well as the presence of a conserved motif in all of these proteins (indicative for a putative muramidase activity) suggest that protein 19 of plasmid R1 facilitates the passage of DNA during conjugation and entry of RNA during phage infection.
MeSH Terms
Amino Acid Sequence
Bacterial Outer Membrane Proteins/biosynthesis,genetics
Base Sequence
Codon, Terminator
Conjugation, Genetic
Conserved Sequence
Escherichia coli/genetics,growth & development,virology
Fimbriae Proteins
Gene Expression Regulation, Bacterial
Molecular Sequence Data
Mutation
Pili, Sex/genetics,metabolism
Plasmids/genetics
RNA Phages/genetics,growth & development,pathogenicity
RNA, Bacterial/biosynthesis,genetics
RNA, Messenger/biosynthesis,genetics
Sequence Homology, Amino Acid
Chemicals
Bacterial Outer Membrane Proteins
Codon, Terminator
RNA, Bacterial
RNA, Messenger
Fimbriae Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Bayer M
Institut für Mikrobiologie, Karl-Franzens-Universität Graz, Austria.
Eferl R
Zellnig G
Teferle K
Dijkstra A
Koraimann G
Högenauer G
References (34)
34 references, click to expand
-
Gene replacement and retrieval with recombinant M13mp bacteriophages.
J Bacteriol. 1989 Jan;171(1):538-46
PMID: 2644206
-
Analysis of the complete nucleotide sequence of the Agrobacterium tumefaciens virB operon.
Nucleic Acids Res. 1988 May 25;16(10):4621-36
PMID: 2837739
-
Genetic and physical map of plasmid NR1: comparison with other IncFII antibiotic resistance plasmids.
Microbiol Rev. 1988 Dec;52(4):433-51
PMID: 3070319
-
Location of the nick at oriT of the F plasmid.
J Mol Biol. 1989 Jun 5;207(3):505-12
PMID: 2668533
-
Nucleotide sequence of the leading region adjacent to the origin of transfer on plasmid F and its conservation among conjugative plasmids.
Mol Gen Genet. 1989 Oct;219(1-2):177-86
PMID: 2693941
-
The sequence of the leading region of the resistance plasmid R1.
Nucleic Acids Res. 1990 Feb 25;18(4):1046
PMID: 2315017
-
A workbench for multiple alignment construction and analysis.
Proteins. 1991;9(3):180-90
PMID: 2006136
-
The sequences of genes bordering oriT in the enterotoxin plasmid P307: comparison with the sequences of plasmids F and R1.
Plasmid. 1990 Sep;24(2):119-31
PMID: 2096398
-
Simultaneous quantification of several mRNA species by solution hybridisation with oligonucleotides.
Nucleic Acids Res. 1991 Jun 25;19(12):3466
PMID: 2062669
-
Murein-metabolizing enzymes from Escherichia coli: sequence analysis and controlled overexpression of the slt gene, which encodes the soluble lytic transglycosylase.
J Bacteriol. 1991 Nov;173(21):6773-82
PMID: 1938883
-
Sequence similarities between the RP4 Tra2 and the Ti VirB region strongly support the conjugation model for T-DNA transfer.
J Biol Chem. 1992 Oct 5;267(28):20471-80
PMID: 1400366
-
Characterization of the Shigella flexneri ipgD and ipgF genes, which are located in the proximal part of the mxi locus.
Infect Immun. 1993 May;61(5):1707-14
PMID: 8478058
-
Subcellular localization of seven VirB proteins of Agrobacterium tumefaciens: implications for the formation of a T-DNA transport structure.
J Bacteriol. 1993 Aug;175(16):5233-41
PMID: 8349563
-
Expression of gene 19 of the conjugative plasmid R1 is controlled by RNase III.
Mol Microbiol. 1993 Aug;9(4):717-27
PMID: 7694035
-
Inactivation of antibiotics and the dissemination of resistance genes.
Science. 1994 Apr 15;264(5157):375-82
PMID: 8153624
-
A conserved domain in putative bacterial and bacteriophage transglycosylases.
Trends Biochem Sci. 1994 Mar;19(3):106-7
PMID: 8203016
-
Genetic complementation analysis of the Agrobacterium tumefaciens virB operon: virB2 through virB11 are essential virulence genes.
J Bacteriol. 1994 Jun;176(12):3646-60
PMID: 8206843
-
Promiscuous DNA transfer system of Agrobacterium tumefaciens: role of the virB operon in sex pilus assembly and synthesis.
Mol Microbiol. 1994 Apr;12(1):17-22
PMID: 7914664
-
Analysis of the sequence and gene products of the transfer region of the F sex factor.
Microbiol Rev. 1994 Jun;58(2):162-210
PMID: 7915817
-
Common ancestry between IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens.
Mol Microbiol. 1994 Nov;14(4):655-68
PMID: 7891554
-
'Holy' proteins. II: The soluble lytic transglycosylase.
Curr Opin Struct Biol. 1994 Dec;4(6):810-3
PMID: 7712284
-
Nucleotide sequence of gene X of antibiotic resistance plasmid R100.
Nucleic Acids Res. 1988 May 25;16(10):4726
PMID: 2837741
-
Mutant drug resistant factors of high transmissibility.
Nature. 1967 May 27;214(5091):885-7
PMID: 6054969
-
A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
Nucleic Acids Res. 1979 Nov 24;7(6):1513-23
PMID: 388356
-
Cloning and polypeptide analysis of the leading region in F plasmid DNA transfer.
Plasmid. 1983 May;9(3):262-72
PMID: 6306705
-
Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
Gene. 1983 Dec;26(1):101-6
PMID: 6323249
-
The F plasmid origin of transfer: DNA sequence of wild-type and mutant origins and location of origin-specific nicks.
EMBO J. 1984 May;3(5):1175-80
PMID: 6329741
-
DNA sequence of the F traALE region that includes the gene for F pilin.
J Bacteriol. 1984 Oct;160(1):395-401
PMID: 6090426
-
Conjugal transfer system of the IncN plasmid pKM101.
J Bacteriol. 1985 Jan;161(1):402-10
PMID: 2857162
-
Rapid and efficient site-specific mutagenesis without phenotypic selection.
Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92
PMID: 3881765
-
Improved plasmid vectors for the isolation of translational lac gene fusions.
Gene. 1984 Nov;31(1-3):269-73
PMID: 6098531
-
Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector.
Gene. 1986;48(1):119-31
PMID: 3549457
-
Surface exclusion genes traS and traT of the F sex factor of Escherichia coli K-12. Determination of the nucleotide sequence and promoter and terminator activities.
J Mol Biol. 1987 Nov 5;198(1):1-11
PMID: 3323526
-
A stable core region of the tra operon mRNA of plasmid R1-19.
Nucleic Acids Res. 1989 Feb 25;17(4):1283-98
PMID: 2564189