Moreno S, Nájera R, Guzmán J, Soberón-Chávez G, Espín G
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62250, México.
J Bacteriol. 1998 May;180(10):2766-9. doi: 10.1128/JB.180.10.2766-2769.1998.
Alginate is essential for encystment in Azotobacter vinelandii. Transcription of the algD gene, which codes for GDP-mannose dehydrogenase, a key enzyme in the alginate biosynthetic pathway, is initiated at two promoters, one of which, p2, has sigmaE consensus sequences. AlgU is the A. vinelandii alternative sigmaE factor. In this study, we constructed an algU mutant (SMU88) which, as expected, is impaired in alginate production, encystment, and transcription of the algD gene from the p2 promoter. Plasmid pJMSAT1, carrying the A. vinelandii algU gene, restored alginate production and encystment to SMU88 and to strain UW136, a naturally occurring algU mutant. Plasmid pSMU865, carrying the A. vinelandii mucABCD genes coding for negative regulators of AlgU activity and previously shown to diminish alginate production in the wild-type strain, ATCC 9046, was shown here to impair encystment and transcription of the algD gene from the p2 algU-dependent promoter. Since nonencysting strain ATCC 9046/pSMU865 produced more alginate than some encysting strains, such as UW136/pJMSAT1, we propose an AlgU role in encystment, independent of the structural role that alginate plays in mature cysts.
藻酸盐对于维氏固氮菌的包囊形成至关重要。编码GDP-甘露糖脱氢酶(藻酸盐生物合成途径中的关键酶)的algD基因的转录起始于两个启动子,其中一个启动子p2具有σE共有序列。AlgU是维氏固氮菌的替代σE因子。在本研究中,我们构建了一个algU突变体(SMU88),正如预期的那样,该突变体在藻酸盐产生、包囊形成以及从p2启动子转录algD基因方面存在缺陷。携带维氏固氮菌algU基因的质粒pJMSAT1恢复了SMU88以及菌株UW136(一种天然存在的algU突变体)的藻酸盐产生和包囊形成能力。携带编码AlgU活性负调节因子的维氏固氮菌mucABCD基因的质粒pSMU865,先前已证明其可减少野生型菌株ATCC 9046中的藻酸盐产生,在此研究中发现它会损害从依赖algU的p2启动子转录algD基因以及包囊形成。由于不形成包囊的菌株ATCC 9046/pSMU865比一些能形成包囊的菌株(如UW136/pJMSAT1)产生更多的藻酸盐,我们提出AlgU在包囊形成中发挥作用,这一作用独立于藻酸盐在成熟包囊中所起的结构作用。