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W Messer

Publications and source records attributed to W Messer.

106 records · Page 6Linked to original sources

Mutants of Escherichia coli B-r defective in deoxyribonucleic acid initiation: dnaI, a new gene for replication.

Mutagenized E. coli B/r cells were subjected to a procedure designed to select mutants temperature-sensitive for initiation of deoxyribonucleic acid (DNA) replication. Seventeen mutants exhibiting limited residual DNA synthesis at 42 C were obtained and the dna(-) sites were mapped genetically. Sixteen of the sites map near dnaA, dnaB, and dnaC. One mutant (dna-208) maps in a new location between the trp and his genes. We propose to call this mutant dnaI208. In complementation experiments dnaC(+) and dnaI(+) were dominant to dnaC(-) and dnaI(-) alleles, respectively. However, dnaA(-) was dominant to the wild-type allele dnaA(+). All dnaA mutants and four out of six dnaC mutants could be suppressed by F factor integration. The pattern of suppression was specific for each mutant.

Bacterial Proteins↗

Initiation of deoxyribonucleic acid replication in Escherichia coli B-r: chronology of events and transcriptional control of initiation.

Three processes necessary for the initiation of deoxyribonucleic acid replication have been separated in time in synchronously growing Escherichia coli B/r. One can be inhibited with 2 mug of chloramphenicol per ml and occurs about 20 min prior to initiation. A second step, occurring 5 to 10 min prior to initiation, is sensitive to 4 to 30 mug of chloramphenicol per ml. A third process occurs at the time of initiation and can be inhibited with rifampin. The experiments suggest that transcription itself, or the synthesis of a relatively small ribonucleic acid, is required for initiation of replication.

Bacterial Proteins↗

Deoxyribonucleic acid replication and cell division in Escherichia coli at 33 C.

At 33 C (60-min generation time) the time required to replicate the chromosome is C = 60 min. The time between the end of a round of replication and cell division is D = 20 min, as at 37 C. Nalidixic acid and a temperature shift in a dnaB mutant give identical results for the determination of the end of a round of replication.

Cell Division↗

The complex of oriC DNA with the DnaA initiator protein.

We describe several experimental approaches relating to the early steps in the initiation of DNA replication at oriC. 1) A matrix is given which enables calculatation of the relative affinity of DnaA boxes for DnaA protein; 2) base changes within single Dna A boxes in oriC have little effect on oriC function; 3) mutations which change the distance between DnaA boxes inactivate oriC, but changes by one helical turn (+ and -) result in near wild-type oriC activity; 4) a Fis binding site was located at oriC coordinates 206-220; 5) KMnO4 probing demonstrates Dna-A-dependent unwinding in the left part of oriC in vivo and in vitro.

Bacterial Proteins↗

Functional domains of DnaA proteins.

Functional domains of the initiator protein DnaA of Escherichia coli have been defined. Domain 1, amino acids 1-86, is involved in oligomerization and in interaction with DnaB. Domain 2, aa 87-134, constitutes a flexible loop. Domain 3, aa 135-373, contains the binding site for ATP or ADP, the ATPase function, a second interaction site with DnaB, and is required for local DNA unwinding. Domain 4 is required and sufficient for specific binding to DNA. We show that there are three different types of cooperative interactions during the DNA binding of DnaA proteins from E. coli, Streptomyces lividans, and Thermus thermophilus: i) binding to distant binding sites; ii) binding to closely spaced binding sites; and iii) binding to non-canonical binding sites.

Amino Acid Sequence↗