1991
DOI: 10.1111/j.1432-1033.1991.tb15775.x
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Isolation and inactivation of the nuclear gene encoding the rotenone‐insensitive internal NADH: ubiquinone oxidoreductase of mitochondria from Saccharomyces cerevisiae

Abstract: We have recently described the isolation of a mitochondrial rotenone-insensitive NADH : ubiquinone oxidoreductase from Saccharomyces cerevisiae [de Vries, S. & Grivell, L. A. (1988) Eur. J. Biochem. 176,3841. We now report the isolation of the nuclear gene encoding this single-subunit enzyme. Null mutants have been constructed by means of one-step gene disruption. Oxygen-uptake experiments, performed with mitochondria isolated from the mutant cells, showed that this NADH dehydrogenase catalyzes the oxidation … Show more

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Cited by 125 publications
(117 citation statements)
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“…There is evidence from complementation studies in yeast and N. crassa that ScNDE1, ScNDE2, and ScNDE1 are NADH-specific enzymes (Marres et al, 1991;Luttik et al, 1998), whereas NcNDE1 is an NADPH-specific enzyme (Melo et al, 2001). From an examination of the NcNDE1 sequence, Melo et al (2001) speculated that the more N-terminal DNF domain binds NADPH as the third Gly is replaced by Ser, and the negative charge at the end of the second ␤-sheet is missing.…”
Section: Substrate Specificity Of Atndi1mentioning
confidence: 99%
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“…There is evidence from complementation studies in yeast and N. crassa that ScNDE1, ScNDE2, and ScNDE1 are NADH-specific enzymes (Marres et al, 1991;Luttik et al, 1998), whereas NcNDE1 is an NADPH-specific enzyme (Melo et al, 2001). From an examination of the NcNDE1 sequence, Melo et al (2001) speculated that the more N-terminal DNF domain binds NADPH as the third Gly is replaced by Ser, and the negative charge at the end of the second ␤-sheet is missing.…”
Section: Substrate Specificity Of Atndi1mentioning
confidence: 99%
“…These have been well characterized (Marres et al, 1991;de Vries et al, 1992;Luttik et al, 1998). Kinetic analyses have also shown these enzyme activities to be present in mitochondria from the fungus Neurospora crassa (Weiss et al, 1970).…”
mentioning
confidence: 98%
“…Consequently, the purified enzyme contained negligible amounts of the proteolytic breakdown product [3]. The higher-molecular-mass band was cut out after SDS/PAGE as described in [5] and used for determination of the N-terminus.…”
Section: Enzyme Purificationmentioning
confidence: 99%
“…coli strains JMlOl [I91 and HBlOl [2O] were used to propagate recombinant DNA constructs. S. cerevisiue strain YPIO2(a, his3,leu2, lysl, ura, ade, NDH) (wild type) was used to obtain the mutant lacking the intact NADH dehydrogenase gene, YP102(a, his3, lysl, ura, ade, NDH : : leu2) [5]. This mutant strain was transformed with the shuttle plasmids YCP5O or YEplacl95 [21] carrying various DNA inserts encoding the internal NADH dehydrogenase.…”
Section: Strains and Growth Conditionsmentioning
confidence: 99%
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