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Biomedical subjects

H Echols

Publications and source records attributed to H Echols.

At least 91 records · Page 5Linked to original sources
PubMed1973

Effect of circularity and superhelicity on transcription from bacteriophagelambda DNA.

We studied RNA synthesis in vitro from closed-circular lambda DNA molecules with varying degrees of superhelicity. The four circular templates examined had 0, -50, -110, and -160 superhelical turns under the conditions of the transcription assay. We found that the total amount of RNA synthesis increases as the template acquires more negative superhelical turns. This increased transcription results from more frequent initiation of RNA chains. Transcription of circular DNA with no superhelical turns appears to mimic RNA synthesis in vivo more closely than transcription from either highly superhelical or linear DNA with regard to two criteria: preferential transcription of the region corresponding to early genes and sensitivity to repression by lambda cI protein. We suggest that the physical basis for the increased initiation of RNA chains from superhelical DNA is the fact that unwinding events are energetically favored on a DNA molecule with negative superhelical turns. Possible general mechanisms are: (a) RNA polymerase must unwind the DNA duplex as a prelude to initiation; (b) the DNA itself must assume a new conformation at the promoter site which requires an unwinding of the DNA duplex.

Centrifugation, Density Gradient↗
PubMed1973

Purification and properties of the gamma-protein specified by bacteriophage lambda: an inhibitor of the host RecBC recombination enzyme.

Previous experiments have indicated that the gam gene of bacteriophage lambda is responsible for an inhibition of the RecBC DNase-an enzyme that is essential for the major host pathway of genetic recombination. We report here experiments that define the inhibitor as the protein product of the gam gene ("gamma-protein") and that characterize the inhibition reaction with highly purified preparations of gamma-protein and RecBC DNase. Genetic characterization was performed with partially purified fractions prepared from cells infected with various lambda mutants. An activity that inhibits RecBC DNase was absent in extracts prepared after infection by phage that carry nonsense or deletion mutations in the gam gene; this activity was highly thermolabile in an extract prepared after infection by phage that carry a temperature-sensitive mutation in the gam gene. For biochemical characterization, the gamma-protein has been purified more than 800-fold. This highly purified preparation inhibited all of the known catalytic activities associated with the RecBC enzyme, but exhibited no detectable DNase or ATPase activities by itself. These findings are discussed in terms of their implications for regulation of genetic recombination and bacteriophage lambda development.

Adenosine Triphosphatases↗
PubMed1971

Establishment and maintenance of repression by bacteriophage lambda: the role of the cI, cII, and c3 proteins.

To define the events necessary for the establishment and maintenance of repression in a lambda-infected cell, we have studied the requirements for efficient synthesis of the cI protein ("lambda-repressor"). Three classes of lambda mutants defective in the establishment of repression are also defective in the appearance of cI protein activity at the normal time. Two of these mutational classes (cII(-) and cIII(-)) probably result from inactivation of lambda-specified proteins, but the third class (cy(-)) may involve a structural defect. We conclude that at least three regulatory elements are likely to be required for the normal turn-on of cI protein synthesis in an infected nonlysogenic cell: cII and cIII proteins and an "active" y-region of lambda DNA. From these and other results, the complete role of cII and cIII proteins in the establishment of repression may involve a bifunctional regulatory activity: positive regulation of the cI gene and negative regulation of late genes. A possible molecular model for cII and cIII action is discussed. Since the cII and cIII genes are repressed by the cI protein under conditions of stable lysogeny, a separate mechanism is required for the maintenance of cI protein synthesis. After infection of a lysogen by cII(-) phage, the rate of increase of cI protein activity is substantially greater than after infection of a nonlysogen. From these and other results, the cI protein may also have a bifunctional regulatory activity: positive regulation of the cI gene and negative regulation of early lytic genes.

Autoradiography↗