Calcium/calmodulin (Ca2+/CaM)-dependent protein kinase II (CaMKII) couples increases in cellular Ca2+ to fundamental responses in excitable cells. CaMKII was identified over 20 years ago by activation dependence on Ca2+/CaM, but recent evidence shows that CaMKII activity is also enhanced by pro-oxidant conditions. Here we show that oxidation of paired regulatory domain methionine residues sustains CaMKII activity in the absence of Ca2+/CaM. CaMKII is activated by angiotensin II (AngII)-induced oxidation, leading to apoptosis in cardiomyocytes both in vitro and in vivo. CaMKII oxidation is reversed by methionine sulfoxide reductase A (MsrA), and MsrA-/- mice show exaggerated CaMKII oxidation and myocardial apoptosis, impaired cardiac function, and increased mortality after myocardial infarction. Our data demonstrate a dynamic mechanism for CaMKII activation by oxidation and highlight the critical importance of oxidation-dependent CaMKII activation to AngII and ischemic myocardial apoptosis.

A statistical thermodynamic model has been developed In prokaryotes, genes are commonly switched on and off by the interactions of regulatory proteins with specific DNA sequences. A particularly complex example of such a switch is found at the right operator (OR) ofbacteriophage A: this operator consists of three tandem DNA sites that are recognized by two phage-encoded regulatory proteins (the A repressor and cro protein). When phage A is in the "lysogenic state," the A repressor (cI gene product) is synthesized and occupies sites OR1 and OR2 In this configuration of OR, the cro gene is repressed and the ci gene is transcribed.The phage switches into the "lytic state" when the repressor protein is cleaved in half by the recA protein, an action initiated by DNA damage. As repressor is destroyed, the cro gene is derepressed and the cro protein is made and occupies site OR3, turning off transcription of the cl gene, yet allowing its own synthesis. In this fashion, the phage can switch from one state (lysogeny) to another (lytic growth) in response to an external signal (for review, see ref. 1).In this paper, we consider the lysogenic state (repressor on, cro off) in an attempt to understand the physical principles that govern the ways in which the protein-DNA and protein-protein interactions operate in concert to produce the known physiological behavior. We show that a model based on statistical thermodynamic assumptions is sufficient to account for some of the known physiological properties of the repressor-operator regulatory system. A brief summary of certain aspects of this work has been presented elsewhere (1).Models for interactions at the lac operon have been developed to include effects of inducer and nonspecific DNA on the binding of lac repressor (2-4). The systems of A and other inducible phages differ from lac by using multiple operator binding sites that have cooperative interactions between bound repressors (1). This requires a more elaborate theoretical approach to the protein-DNA binding problem-one that has not previously been developed. The mathematical model we present here incorporates a set of rules and assumptions derived from previous genetic, biochemical, and structural studies. Combination of this information with statistical thermodynamic assumptions generates a quantitative formulation that incorporates salient features of the qualitative description developed over the last several years (1,(5)(6)(7)(8)(9)). This quantitative model has the following significance. (i) It provides a way to test assumptions regarding the physical bases for operation of the system. Thus, it predicts quantitatively the activities of the two OR-controlled promoters (PR and PRM) as a function of repressor concentration, and these predictions can then be compared with known.physiological properties of the system. (ii) It points out features of the A operator system that were not obvious (or at least fully appreciated) from previous experiments. (iii) It incorporates a theory for interpreting cooperati...

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