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. 1976 Aug;127(2):770–779. doi: 10.1128/jb.127.2.770-779.1976

Regulation of molybdate transport by Clostridium pasteurianum.

B B Elliott, L E Mortenson
PMCID: PMC232983  PMID: 956118

Abstract

The regulation of the molybdate (MoO42-) transport activity of Clostridium pasteurianum has been studied by observing the effects of NH3, carbamyl phosphate, MoO42-, and chloramphenicol on the ability of cells to take up MoO42-. Compared with cells fixing N2, cells grown in the presence of 1 mM NH3 are greater than 95% repressed for MoO42- transport. Uptake activity begins to increase just before NH exhaustion (under Ar or N2) and continues to increase throughout the lag period as cells shift from NH3-growing to N2-fixing conditions. When cells are shifted from N2-fixing to NH3-growing conditions the transport activity per fixed number of cells decreases by increase of bells in absence of transport synthesis. Carbamyl phosphate (greater than or equal to 15 mM) but not NH3 inhibits 58% of the in vitro uptake activity. When 1 mM carbamyl phosphate is added just before the exhaustion of NH3, the transport activity, measured 2 h later, is 100% repressed. Cells grown in the presence of high MoO42- (1mM) are 80% repressed for MoO42- transport. Synthesis of the MoO42- transport system is also completely stopped when chloramphenicol (300 mug/ml) is added just before the exhaustion oNH 3 from the medium. These findings demonstrate that the ability of cells to transport MoO42- is dependent upon new protein synthesis and can be repressed by high levels of substrate. The regulation of MoO42- uptake by NH3 or carbamyl phosphate closely parallels the regulation of nitrogenase activity. Activity of neither nitrogenase component (Fe protein or MoFe protein) was detected even 3 h after the exhaustion of the NH3 if either MoO42- was absent or if WO42- was present in place of MoO42-. The duration of the diauxic lag increases with decreasing concentration of MoO42- in the medium. If no MoO42- is present the lag continues indefinitely. If MoO42- is added late in the lag period, growth under N2-fixing conditions resumes but only after a normal induction period.

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Selected References

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  1. Benemann J. R., Smith G. M., Kostel P. J., McKenna C. E. Tungsten incorporation into Azotobacter vinelandii nitrogenase. FEBS Lett. 1973 Feb 1;29(3):219–221. doi: 10.1016/0014-5793(73)80023-7. [DOI] [PubMed] [Google Scholar]
  2. Brill W. J., Steiner A. L., Shah V. K. Effect of molybdenum starvation and tungsten on the synthesis of nitrogenase components in Klebsiella pneumonia. J Bacteriol. 1974 Jun;118(3):986–989. doi: 10.1128/jb.118.3.986-989.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bulen W. A., LeComte J. R. The nitrogenase system from Azotobacter: two-enzyme requirement for N2 reduction, ATP-dependent H2 evolution, and ATP hydrolysis. Proc Natl Acad Sci U S A. 1966 Sep;56(3):979–986. doi: 10.1073/pnas.56.3.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cardenas J., Mortenson L. E. Determination of molybdenum and tungsten in biological materials. Anal Biochem. 1974 Aug;60(2):372–381. doi: 10.1016/0003-2697(74)90244-9. [DOI] [PubMed] [Google Scholar]
  5. Cardenas J., Mortenson L. E. Role of molybdenum in dinitrogen fixation by Clostridium pasteurianum. J Bacteriol. 1975 Sep;123(3):978–984. doi: 10.1128/jb.123.3.978-984.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daesch G., Mortenson L. E. Effect of ammonia on the synthesis and function of the N 2 -fixing enzyme system in Clostridium pasteurianum. J Bacteriol. 1972 Apr;110(1):103–109. doi: 10.1128/jb.110.1.103-109.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eady R. R., Smith B. E., Cook K. A., Postgate J. R. Nitrogenase of Klebsiella pneumoniae. Purification and properties of the component proteins. Biochem J. 1972 Jul;128(3):655–675. doi: 10.1042/bj1280655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Elliott B. B., Mortenson L. E. Transport of molybdate by Clostridium pasteurianum. J Bacteriol. 1975 Dec;124(3):1295–1301. doi: 10.1128/jb.124.3.1295-1301.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Mortenson L. E., Morris J. A., Jeng D. Y. Purification, metal composition and properties of molybdoferredoxin and azoferredoxin, two of the components of the nitrogen-fixing system of Clostridium pasteurianum. Biochim Biophys Acta. 1967 Aug 29;141(3):516–522. doi: 10.1016/0304-4165(67)90180-8. [DOI] [PubMed] [Google Scholar]
  11. Seto B. L., Mortenson L. E. Mechanism of carbamyl phosphate inhibition of nitrogenase of Clostridium pasteurianum. J Bacteriol. 1974 Feb;117(2):805–812. doi: 10.1128/jb.117.2.805-812.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Seto B., Mortenson L. E. Effect of carbamyl phosphate on the regulation of nitrogenase in Clostridium pasteurianum. Biochem Biophys Res Commun. 1973 Jul 17;53(2):419–423. doi: 10.1016/0006-291x(73)90678-5. [DOI] [PubMed] [Google Scholar]
  13. Seto B., Mortenson L. E. In vivo kinetics of nitrogenase formation in Clostridium pasteurianum. J Bacteriol. 1974 Nov;120(2):822–830. doi: 10.1128/jb.120.2.822-830.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Zelitch I. Simultaneous Use of Molecular Nitrogen and Ammonia by Clostridium Pasteurianum. Proc Natl Acad Sci U S A. 1951 Sep;37(9):559–565. doi: 10.1073/pnas.37.9.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Zumft W. G., Mortensson L. E. Evidence for a catalytic-centre heterogeneity of molybdoferredoxin from Clostridium pasteurianum. Eur J Biochem. 1973 Jun 15;35(3):401–409. doi: 10.1111/j.1432-1033.1973.tb02852.x. [DOI] [PubMed] [Google Scholar]

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