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• W2500565562 abstract "Judith W. Zyskind’ and Douglas W. Smith1 *Department of Biology San Diego State University San Diego, California 92182 rDepartment of Biology, 0322 and the Center for Molecular Genetics University of California, San Diego La Jolla, California 92093 Three cellular processes occur during conversion of a mother bacterial cell into daughter cells: cell growth, DNA replication, and cell division. These processes are cou- pled, but the mechanisms that coordinate them are un- known. Bacterial cells rarely make errors in these pro- cesses during their cell cycle-less than 0.03% of the daughter cells contain no DNA. In prokaryotic cells, the rate of replication is controlled not by the rate of chain elongation, but by the rate of initiation at the origin of repli- cation, 06. Thus, the coupling of replication to cell growth requires that the frequency of initiation from oriC be a func- tion of growth rate. Our understanding of the bacterial cell cycle (see figure) is based on the classic work of Cooper and Helmstetter (for review, see Helmstetter, 1987). The time in the cell cycle dedicated to replication (C) is the time between initia- tion (i) and termination (t) of a round of replication. Except in slow-growing cells, the C period (approximately 42 min) varies only slightly with growth rate. D, which is the time between termination (t) and cell division (d), is also invari- ant with growth rate (22-25 min). If the sum of C and D is less than the doubling time (7) agap between cell division and initi’ation (B) occurs (part A of figure). If, however, the sum of C and D is greater than T, replication reinitiates before completion of the previous round of replication (part 6 of figure), leading to overlapping C+D periods in the same cell. For T = 27 min (part B of figure), newborn cells contain four origins (0) and one terminus(T) of replication. Two replication forks (f2) are assembled at time i2, and four of the replication forks (f,) are assembled at time i3. Independent of the length C+D, critical period in coordinating replication with cell growth is the interinitia- tion time, which is always equal to the doubling time 5. This review discusses three ways that the length of interini- tiation time is likely to be regulated during cell growth; two regulate the time at which initiation events occur in the cell cycle, and a third responds to changes in cell growth. initiation of Replication Is Coupled to the Celf Cycle by the Concentration of DnaA The concentration of DnaA is critical in determining the timing of initiation, the first step of which is the formation of a complex of 20-40 molecules of DnaA with oriC (Fun- nell et al., 1987). This interaction involves binding to four repeats called dnaA boxes, for which a new consensus sequence has been derived (Schaefer and Messer, 1991). DnaA then opens the DNA helix at AT-rich 13-mer repeats in oriC, prior to loading of DnaB helicase by DnaC (Bramhill and Kornberg, 1988). Increased synthesis of DnaA stimulates initiation of replication. When dnaA ex- pression is induced, the number origins and DNA content per mass unit increase (Skarstad et al., 1989; Lebner-Olesen et al., 1989). De novo protein synthesis is required for reinitiation. Experimental results support the possibility that it is at least additional DnaA that must be synthesized. Blockage of additional DnaA synthesis in cfnaA amber mutants by heat inactivation of a temperature-sensitive suppressor tRNAcausescessation of DNA accumulation and progres- sive inhibition of the rate of DNA synthesis. The kinetics DNA pulse-labeling in these experiments are those pre- dicted for inhibition of initiation with completion of existing rounds of replication (Schaus et al., 1981). These results indicate either that there is insufficient DnaA for another initiation event or that DnaA, once having" @default.
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• W2500565562 date "1992-01-01" @default.
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• W2500565562 title "DNA Replication, the Bacterial Cell Cycle, and Cell Growth Minireview" @default.
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