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S Kitayama

Publications and source records attributed to S Kitayama.

At least 127 records · Page 7Linked to original sources

An exonuclease activity associated with DNA polymerase I of Micrococcus radiodurans.

An exonuclease activity is associated with one of three DNA polymerase in Micrococcus radiodurans. The nuclease activity co-sedimented with its DNA polymerase I of this bacterium on glycerol gradient centrifugation. Both activities show the same optimum pH and heat-inactivation kinetics. This nuclease hydrolyzes preferentially double-stranded DNA in an exonucleolytic manner from both ends of the duplex DNA. The products of hydrolysis are mostly deoxyribonucleoside 5'-monophosphate and no nucleosides are released into the acid-soluble fraction. Di- or other oligonucleotides are also produced but their relative amounts are constant during the time of incubation. The exonuclease activity requires Mg2+ and is inhibited by high concentrations of KCl as is DNA polymerase I of M. radiodurans.

DNA Polymerase I↗

Separation of DNA-dependent DNA polymerase activities in Micrococcus radiodurans.

DNA polymerase activities in Micrococcus radiodurans were separated into two fractions after purification more than 2000 fold. They differ in pH optimum and residual activities in the absence of a full deoxyribonucleoside triphosphates complement. NAD partly inhibited one of the activities. Both activities were eluted as a single peak on gel filtration and sedimented at the same rate on glycerol gradient centrifugation. Molecular weight 140000 was calculated from Stokes radius and sedimentation constant. Deoxyribonuclease activity was detected on one of the polymerase activities which preferentially degraded double-stranded DNA. Priming activity of nicked DNA was reduced by gamma-irradiation. These results have been related to the possible rolls in repair synthesis in vivo or DNA synthesis in permeable cells of M. radiodurans.

DNA↗

DNA synthesis and repair in permeable cells of Micrococcus radiodurans.

Cells permeable to deoxyribonucleoside triphosphate were prepared from Micrococcus radiodurans, and DNA synthesis and rejoining of strand scissions induced by gamma-rays were investigated. DNA synthesis was stimulated by ATP at an optimal concentration of 1mM. This reaction requires four deoxyribonucleoside triphosphates and MgCl2. NAD inhibited the reaction, but no rejoining of primer DNA was observed. Even in the presence of NAD, DNA which was synthesized in the unirradiated permeable cells had a peak molecular weight of only 1.3 - 10(6). DNA synthesis was stimulated by irradiation of the permeable cells with gamma-rays, but this stimulatory effect was eliminated by the addition of NAD. Both primer and synthesized DNA in the irradiated permeable cells were rejoined in vitro in the presence of NAD and deoxyribonucleoside triphosphates, while those in the unirradiated permeable cells were not rejoined.

Biological Transport↗