Bradley S Nefsky

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Ded1 is a fission yeast DEAD box protein involved in translation. We isolated Ded1 in a screen for multi-copy suppressors of a cold-sensitive, loss-of-function mutant of the cyclin-dependent kinase Cdc2. The checkpoint protein kinase Chk1, required for cell cycle arrest in response to DNA damage, was also isolated in this screen. Ded1 interacts with Chk1 in(More)
Caldesmons are major Ca2+-calmodulin regulated F-actin binding proteins of smooth and non-muscle cells that have been implicated as components of a thin filament regulatory system. Chicken gizzard caldesmons are monomeric proteins of Mr 140,000 and 135,000. We have employed enzymatic and chemical cleavage methods in order to dissect the protein to locate(More)
Although the molecular mechanism of aging is unknown, a progressive increase with age in the concentration of damaged macromolecules, especially proteins, is likely to play a central role in senescent decline. In this paper, we discuss evidence that the progressive decrease in protein synthesis and turnover can be the primary cause of the increase in the(More)
Actin from yeast has been reported previously to have unusual polymerization properties. Here we report a simple sensitive spot assay for actin and use it to develop a high-yield procedure for the purification of actin from the yeast Saccharomyces cerevisiae. The polymerization properties of purified yeast actin are quantitatively similar to all other(More)
We describe a general method to locate the positions of cysteine residues relative to the amino terminus of a protein, using a modified chemical cleavage of the polypeptide backbone at cysteine. The cleavage reaction introduces the carbon atom of 14CN into the carboxyl-terminal fragment produced at each cleavage of the polypeptide chain. Peptides containing(More)
A procedure is described for the immobilization of monomeric actin so that about 30% of the immobilized protein is competent to bind the monomeric-actin-binding proteins bovine pancreatic deoxyribonuclease I and chicken villin. The intact tertiary structure of the immobilized actin is required to bind these proteins. Using this resin, a method has been(More)
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