In situ activity of enzymes on polyacrylamide gels of a deoxyribonucleic acid-membrane fraction extracted from pneumococci.
A deoxyribonucleic acid (DNA)-membrane fraction extracted from Diplococcus pneumoniae was subjected to polyacrylamide gel electrophoresis after treatment with 0.16% sodium dodecyl sulfate. At least two DNA polymerase activities were detected by in situ assays with appropriate substrates, templates, and inhibitors, including a co-polymer of deoxyadenylic and thymidylic acid and N-ethylmaleimide. This activity coincided with a fraction in the gel containing 7.5, 9.4, and 24%, respectively of the DNA, phospholipid, and protein present in the DNA-membrane fraction before electrophoresis and sodium dodecyl sulfate treatment. Assays with minced gels showed that several nuclease activities, deoxyribonucleotide kinase activity, and DNA ligase activity also coincided with this fraction. However, ribonucleoside diphosphate reductase activity did not. These results demonstrate that a complex of enzymes involved in DNA replication is firmly bound to the DNA-membrane fraction in pneumococci.
D001426:Bacterial Proteins / Q000032:analysis; D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000032:analysis / Q000201:enzymology*; D002474:Cell-Free System; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D004261:DNA Replication; D004269:DNA, Bacterial / Q000032:analysis / Q000096:biosynthesis*; D003851:Deoxyribonucleases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008025:Ligases / Q000378:metabolism; D008722:Methods; D009097:Multienzyme Complexes / Q000378:metabolism*; D009691:Nucleic Acid Denaturation; D010743:Phospholipids / Q000032:analysis; D010770:Phosphotransferases / Q000378:metabolism; D012264:Ribonucleotide Reductases / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D013296:Streptococcus pneumoniae / Q000032:analysis / Q000201:enzymology* / Q000378:metabolism
D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D004269:DNA, Bacterial; D009097:Multienzyme Complexes; D010743:Phospholipids; D012967:Sodium Dodecyl Sulfate; D012264:Ribonucleotide Reductases; D010770:Phosphotransferases; D004254:DNA Nucleotidyltransferases; D003851:Deoxyribonucleases; D008025:Ligases