Núñez C, Moreno S, Soberón-Chávez G, Espín G
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.
J Bacteriol. 1999 Jan;181(1):141-8. doi: 10.1128/JB.181.1.141-148.1999.
Azotobacter vinelandii produces the exopolysaccharide alginate, which is essential for the encystment process. In Pseudomonas aeruginosa, as well as in A. vinelandii, the sigmaE factor encoded by algU is required for transcription of algD, which encodes a key enzyme of the alginate biosynthetic pathway. The P. aeruginosa response regulator AlgR activates transcription of algD. fimS, located upstream algR, is proposed to encode the AlgR cognate sensor kinase. We have cloned and characterized the A. vinelandii algR gene; the deduced amino acid sequence of the protein encoded by this gene shows 79% identity with its P. aeruginosa homolog. Sequence analysis around the algR gene revealed the absence of a fimS homolog. Inactivation of A. vinelandii algR diminished alginate production by 50%, but did not affect algD transcription, and completely impaired the capacity to form mature cysts. Electron microscopy of the cyst structures formed by the algR mutant revealed that the encystment process is blocked at the step of exine formation. The transcriptional regulation of the A. vinelandii algR gene and the role of AlgR in alginate production differ significantly from those of its P. aeruginosa counterparts. These differences could be due to the fact that in A. vinelandii, alginate plays a role in encystment, a function not found in P. aeruginosa.
棕色固氮菌产生胞外多糖藻酸盐,这对于包囊形成过程至关重要。在铜绿假单胞菌以及棕色固氮菌中,algU编码的σE因子是algD转录所必需的,algD编码藻酸盐生物合成途径的一种关键酶。铜绿假单胞菌应答调节因子AlgR激活algD的转录。位于algR上游的fimS被认为编码AlgR同源感应激酶。我们已经克隆并鉴定了棕色固氮菌的algR基因;该基因编码的蛋白质的推导氨基酸序列与其铜绿假单胞菌同源物显示出79%的同一性。algR基因周围的序列分析表明不存在fimS同源物。棕色固氮菌algR的失活使藻酸盐产量降低了50%,但不影响algD转录,并且完全损害了形成成熟包囊的能力。对algR突变体形成的包囊结构进行电子显微镜观察发现,包囊形成过程在外壁形成步骤受阻。棕色固氮菌algR基因的转录调控以及AlgR在藻酸盐产生中的作用与其铜绿假单胞菌对应物有显著差异。这些差异可能是由于在棕色固氮菌中,藻酸盐在包囊形成中起作用,而在铜绿假单胞菌中未发现这种功能。