The day/night switch of the circadian clock of synechococcus elongatus and hydrogen bonds of dna and rna
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The circadian oscillator of the cyanobacterium Synechococcus elongatus is composed of only three proteins, KaiA, KaiB, and KaiC, which together with ATP can generate a self-sustained ~24 hour oscillation of KaiC phosphorylation for several days. KaiA induces KaiC to autophosphorylate whereas KaiB blocks the stimulation of KaiC by KaiA, which allows KaiC to autodephosphorylate. We propose and support a model in which the C-terminal loops of KaiC, the “A-loops”, are the master switch that determines overall KaiC activity. When the A-loops are in their buried state, KaiC is an autophosphatase. When the A-loops are exposed, however, KaiC is an autokinase. The data suggest that KaiA stabilizes the exposed state of the A-loops through direct binding. We also show evidence that if KaiA cannot stabilize the exposed state KaiC remains hypophosphorylated. We propose that KaiB inactivates KaiA by preventing it from stabilizing the exposed state of the A-loops. Thus, KaiA and KaiB likely act by shifting the dynamic equilibrium of the A-loops between exposed and buried states, which shifts the balance of autokinase and autophosphatase activities of KaiC. A-loop exposure likely moves the ATP closer to the sites of phosphorylation and we show evidence in support of how this movement may be accomplished. Density functional theory calculations of isolated Watson–Crick A:U and A:T base pairs predict that adenine 13C2 trans-hydrogen bond deuterium isotope shifts due to isotopic substitution at the pyrimidine H3, 2hΔ13C2, are sensitive to the hydrogen-bond distance between the N1 of adenine and the N3 of uracil or thymine, which supports the notion that 2hΔ13C2 is sensitive to hydrogen-bond strength. Calculated 2hΔ13C2 values at a given N1–N3 distance are the same for isolated A:U and A:T base pairs. Replacing uridine residues in RNA with 5-methyl uridine and substituting deoxythymidines in DNA with deoxyuridines do not statistically shift empirical 2hΔ13C2 values. Thus, we show experimentally and computationally that the C7 methyl group of thymine has no measurable affect on 2hΔ13C2 values. Furthermore, 2hΔ13C2 values of modified and unmodified RNA are more negative than those of modified and unmodified DNA, which supports our hypothesis that RNA hydrogen bonds are stronger than those of DNA. It is also shown here that 2hΔ13C2 is context dependent and that this dependence is similar for RNA and DNA.
Kim, Yong-Ick (2008). The day/night switch of the circadian clock of synechococcus elongatus and hydrogen bonds of dna and rna. Doctoral dissertation, Texas A&M University. Available electronically from