Kim K S, Farrand S K
Department of Microbiology, University of Illinois at Urbana-Champaign, 61801, USA.
J Bacteriol. 1996 Jun;178(11):3275-84. doi: 10.1128/jb.178.11.3275-3284.1996.
Agrobacterium tumefaciens NT1 harboring pSaB4, which contains the 14-kb BamHI fragment 4 from the octopine/mannityl opine-type Ti plasmid pTi15955, grew well with agropine (AGR) but slowly with mannopine (MOP) as the sole carbon source. When a second plasmid encoding a dedicated transport system for MOP was introduced, these cells grew well with both AGR and MOP. Transposon insertion mutagenesis and subcloning identified a 5.7-kb region of BamHI fragment 4 that encodes functions required for the degradation of MOP. DNA sequence analysis revealed seven putative genes in this region: mocD (moc for mannityl opine catabolism) and mocE, oriented from right to left, and mocRCBAS, oriented from left to right. Significant identities exist at the nucleotide and derived amino acid sequence levels between these moc genes and the mas genes that are responsible for opine biosynthesis in crown gall tumors. MocD is a homolog of Mas2, the anabolic conjugase encoded by mas2'. MocE and MocC are related to the amino half and the carboxyl half, respectively, of Mas1 (MOP reductase), the second enzyme for MOP biosynthesis. These results indicate that the moc and mas genes evolved from a common origin. MocR and MocS are related to each other and to a putative repressor for the AGR degradation system encoded by the rhizogenic plasmid pRiA4. MocB and MocA are homologs of 6-phosphogluconate dehydratase and glucose-6-phosphate dehydrogenase, respectively. Mutations in mocD and mocE, but not mocC, are suppressed by functions encoded by the chromosome or the 450-kb megaplasmid present in many Agrobacterium isolates. We propose that moc genes derived from genes located elsewhere in the bacterial genome and that the tumor-expressed mas genes evolved from the bacterial moc genes.
携带pSaB4的根癌农杆菌NT1,其含有来自章鱼碱/甘露碱型Ti质粒pTi15955的14kb BamHI片段4,以农杆碱(AGR)为唯一碳源时生长良好,而以甘露碱(MOP)为唯一碳源时生长缓慢。当引入编码专门用于MOP转运系统的第二个质粒时,这些细胞在AGR和MOP两种碳源上都生长良好。转座子插入诱变和亚克隆鉴定出BamHI片段4的一个5.7kb区域,该区域编码MOP降解所需的功能。DNA序列分析揭示了该区域的七个推定基因:从右到左排列的mocD(甘露碱分解代谢的moc)和mocE,以及从左到右排列的mocRCBAS。这些moc基因与冠瘿瘤中负责章鱼碱生物合成的mas基因在核苷酸和推导的氨基酸序列水平上存在显著的一致性。MocD是由mas2'编码的合成共轭酶Mas2的同源物。MocE和MocC分别与MOP生物合成的第二种酶Mas1(MOP还原酶)的氨基端和羧基端相关。这些结果表明moc和mas基因起源于共同的祖先。MocR和MocS相互关联,并且与发根质粒pRiA4编码的AGR降解系统的推定阻遏物相关。MocB和MocA分别是6-磷酸葡萄糖酸脱水酶和6-磷酸葡萄糖脱氢酶的同源物。mocD和mocE的突变,但不是mocC的突变,被许多农杆菌分离株中染色体或450kb大质粒编码的功能所抑制。我们提出moc基因源自细菌基因组其他位置的基因,并且肿瘤中表达的mas基因从细菌的moc基因进化而来。