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1.
J Bacteriol ; 176(8): 2143-50, 1994 Apr.
Article in English | MEDLINE | ID: mdl-8157582

ABSTRACT

We isolated and characterized mutants defective in nuo, encoding NADH dehydrogenase I, the multisubunit complex homologous to eucaryotic mitochondrial complex I. By Southern hybridization and/or sequence analysis, we characterized three distinct mutations: a polar insertion designated nuoG::Tn10-1, a nonpolar insertion designated nuoF::Km-1, and a large deletion designated delta(nuoFGHIJKL)-1. Cells carrying any of these three mutations exhibited identical phenotypes. Each mutant exhibited reduced NADH oxidase activity, grew poorly on minimal salts medium containing acetate as the sole carbon source, and failed to produce the inner, L-aspartate chemotactic band on tryptone swarm plates. During exponential growth in tryptone broth, nuo mutants grew as rapidly as wild-type cells and excreted similar amounts of acetate into the medium. As they began the transition to stationary phase, in contrast to wild-type cells, the mutant cells abruptly slowed their growth and continued to excrete acetate. The growth defect was entirely suppressed by L-serine or D-pyruvate, partially suppressed by alpha-ketoglutarate or acetate, and not suppressed by L-aspartate or L-glutamate. We extended these studies, analyzing the sequential consumption of amino acids by both wild-type and nuo mutant cells growing in tryptone broth. During the lag and exponential phases, both wild-type and mutant cells consumed, in order, L-serine and L-aspartate. As they began the transition to stationary phase, both cell types consumed L-tryptophan. Whereas wild-type cells then consumed L-glutamate, glycine, L-threonine, and L-alanine, mutant cells utilized these amino acids poorly. We propose that cells defective for NADH dehydrogenase I exhibit all these phenotypes, because large NADH/NAD+ ratios inhibit certain tricarboxylic acid cycle enzymes, e.g., citrate synthase and malate dehydrogenase.


Subject(s)
Escherichia coli/enzymology , Genes, Bacterial/physiology , Mutation/physiology , NADH, NADPH Oxidoreductases/metabolism , Amino Acid Sequence , Amino Acids/metabolism , Base Sequence , Electron Transport Complex I , Escherichia coli/genetics , Escherichia coli/growth & development , Genes, Bacterial/genetics , Molecular Sequence Data , NADH, NADPH Oxidoreductases/genetics , Phenotype
3.
Cancer ; 41(3): 1137-9, 1978 Mar.
Article in English | MEDLINE | ID: mdl-638957

ABSTRACT

The results of the 25+ year Cancer Detection Center study, including 20,000 participants and 100,000+ patient-years experience, demonstrate the obviation of appearance of most lower bowel cancers associated with a program of proctosigmoidoscopy and adenomatous polyp removal. Once on a schedule of periodic examinations, study participants developed only a fraction of the anticipated number of rectal cancers; most of the cancers which did develop were detected while at an early stage and with involvement of only the mucosa. Not a single death from lower bowel cancer has occurred among active study participants or for 7+ years following the most recent examination at the Center.


Subject(s)
Precancerous Conditions/diagnosis , Rectal Neoplasms/prevention & control , Adenocarcinoma/diagnosis , Adenocarcinoma/surgery , Female , Humans , Male , Precancerous Conditions/surgery , Rectal Neoplasms/diagnosis , Rectal Neoplasms/surgery , Sigmoidoscopy
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