Differential organization and transcription of the cat2 gene cluster in aniline-assimilating Acinetobacter lwoffii K24.
CatABC genes encode proteins that are responsible for the first three steps of one branch of the beta-ketoadipate pathway involved in the degradation of various aromatic compound by bacteria. Aniline-assimilating Acinetobacter lwoffii K24 is known to have the two-catABC gene clusters (cat1 and cat2) on the chromosome (Kim et al., J. Bacteriol., 179: 5226-5231, 1997). The order of the cat2 gene cluster is catB2A2C2, which has not been found in other bacteria. In this report, we analyzed the transcriptional pattern of the cat2 gene cluster and completely sequenced a 5.8 kbp fragment containing the compactly clustered catB2A2C2 genes and four ORFs. Similar to the ORF(R1) of the cat1 gene cluster, an ORF highly homologous with the catR gene was found 102 by upstream of the catB2 gene and was designated as ORF(R2). Three ORFs, one putative reductase component (ORF(X2)) and two putative LysR family regulatory proteins (ORF(Y2), ORF(Z2)) were located next to the catC2 gene in the opposite direction of the cat2 gene cluster. Two ORFs, ORF(X2) and ORF(Y2), were significantly homologous with tdnB and tdnR of the aniline oxygenase complex of Pseudomonas putida UCC22. RT-PCR analysis and Northern blotting revealed that the catB2 gene is independently transcribed and that the catA2C2 genes are cotranscribed. A primer extension assay revealed that transcription of the catA2C2 gene starts in the C-terminal region of the catB2 gene. These results suggest that the cat2 gene cluster may be under a different gene adaptation from other cat gene clusters.
Kim
S I
SI
Biomolecule Research Group, Korea Basic Science Institute, Taejon 305-333, Korea.
Ha
K S
KS
Leem
S H
SH