TAILIEUCHUNG - Báo cáo Y học: The DNA-polymerase inhibiting activity of poly(b-L-malic acid) in nuclear extract during the cell cycle of Physarum polycephalum

The naturally synchronous plasmodia of myxomycetes synthesize poly(b-L-malic acid), which carries out cell-specific functions. In Physarum polycephalum, poly(b-L-malate) [the salt form of poly(b-L-malic acid)] is highly concentrated in the nuclei, repressing DNA synthetic activity of DNA polymerases by the formation of reversible complexes. To test whether this inhibitory activity is cell-cycle-dependent, purified DNA polymerase a of P. polycephalum was added to the nuclear extract and the activity was measured by the incorporation of [3H]thymidine 5¢-monophosphate into acid precipitable nick-activated salmon testis DNA. Maximum DNA synthesis by the reporter was measured in S-phase, equivalent to a minimum of inhibitory activity | Eur. J. Biochem. 269 1253-1258 2002 FEBS 2002 The DNA-polymerase inhibiting activity of poly b-L-malic acid in nuclear extract during the cell cycle of Physarum polycephalum Sabine Doerhoefer1 Christina Windisch1 Bernhard Anaerer1 Olaa I. Lavrik2. Bona-Seon Lee1 and Eggehard Holler1 1Institut fur Biophysik undphysikalische Biochemie Universitdt Regensburg Germany Novosibirsk Institute of Biorganic Chemistry Siberian Division of the Russian Academy of Sciences Novosibirsk Russia The naturally synchronous plasmodia of myxomycetes synthesize poly b-L-malic acid which carries out cell-spe-cilic functions. In Physarum polycephalum poly b-L-malate the salt form of poly b-L-malic acid is highly concentrated in the nuclei repressing DNA synthetic activity of DNA polymerases by the formation of reversible complexes. To test whether this inhibitory activity is cell-cycle-dependent purihed DNA polymerase a of P. polycephalum was added to the nuclear extract and the activity was measured by the incorporation of 3H thymidine 5 -monophosphate into acid precipitable nick-activated salmon testis DNA. Maximum DNA synthesis by the reporter was measured in S-phase equivalent to a minimum of inhibitory activity. To test for the activity of endogenous DNA polymerases DNA synthesis was followed by the highly sensitive photoaffinity labeling technique. Labeling was observed in S-phase in agreement with the minimum of the inhibitory activity. The activity was constant throughout the cell cycle when the inhibition was neutralized by the addition of spermidine hydrochloride. Also the concentration of poly b-L-malate did not vary with the phase of the cell cycle Schmidt A. Windisch C. Holler E. 1996 Nuclear accumulation and homeostasis of the unusual polymer poly b-L-malate in plasmodia of Physarum polycephalum. Eur. J. Cell Biol. 70 373-380 . To explain the variation in the cell cycle a periodic competition for poly b-L-malate between DNA polymerases and most likely certain histones was .

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