Genetic and functional analysis of the basic replicon of pPS10, a plasmid specific for Pseudomonas isolated from Pseudomonas syringae patovar savastanoi.
The sequence of a 1823 base-pair region containing the replication functions of pPS10, a narrow host-range plasmid isolated from a strain of Pseudomonas savastanoi, is reported. The origin of replication, oriV, or pPS10 is contained in a 535 base-pair fragment of this sequence that can replicate in the presence of trans-acting function(s) of the plasmid. oriV contains four iterons of 22 base-pairs that are preceded by G+C-rich and A+T-rich regions. A dnaA box located adjacent to the repeats of the origin is dispensable but required for efficient replication of pPS10; A and T are equivalent bases at the 5' end of the box. repA, the gene of a trans-acting replication protein of 26,700 Mr has been identified by genetic and functional analysis. repA is adjacent to the origin of replication and is preceded by the consensus sequences of a typical sigma 70 promoter of Escherichia coli. The RepA protein has been identified, using the minicell system of E. coli, as a polypeptide with an apparent molecular mass of 26,000. A minimal pPS10 replicon has been defined to a continuous 1267 base-pair region of pPS10 that includes the oriV and repA sequences.
Nieto
C
C
Centro de Investigaciones Biológicas, C.S.I.C., Madrid, Spain.
Giraldo
R
R
Fernández-Tresguerres
E
E
Díaz
R
R
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D004265:DNA Helicases; D004261:DNA Replication / Q000235:genetics; D004268:DNA-Binding Proteins; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000235:genetics*; D011487:Protein Conformation; D011506:Proteins; D011549:Pseudomonas / Q000235:genetics*; D016364:Reading Frames; D011993:Recombinant Fusion Proteins; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012093:Replicon / Q000235:genetics*; D015534:Trans-Activators
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C066312:DnaA protein, Bacteria; D011506:Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004265:DNA Helicases