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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 259 (1998), S. 429-436 
    ISSN: 1617-4623
    Keywords: Key words Protein phosphorylation ; Allosteric regulation ; DNA replication ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cdc7/Dbf4 protein kinase is required for the initiation of DNA replication in Saccharomyces cerevisiae. Cdc7/Dbf4 protein kinase is not a cyclin-dependent kinase (CDK), but is regulated in a similar fashion in that the Cdc7 kinase subunit is inactive in the absence of the regulatory subunit Dbf4. In contrast to what is known about CDKs, Cdc7/Dbf4 protein kinase is shown to be an oligomer in the cell in this report. Genetic data that support this claim include interallelic complementation between several cdc7ts alleles and the cdc7T281A allele and also the results of experiments using the two-hybrid system with Cdc7 in both DNA-binding and transactivation domain plasmids. A molecular interaction between two different Cdc7 molecules was shown by using a HA-tagged Cdc7 protein that differs in size from the wild-type Cdc7 protein: an anti-HA antibody immunoprecipitates both proteins in appproximately equal stoichiometry. Analysis of the native molecular weight of Cdc7/Dbf4 protein kinase is consistent with oligomerization of the Cdc7 protein in that complexes of about 180 and 300 kDa were found. Oligomers of Cdc7 protein may exist for the purpose of allosteric regulation or to allow phosphorylation of multiple substrate protein molecules.
    Type of Medium: Electronic Resource
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  • 2
    Publication Date: 2016-02-20
    Description: There is widespread agreement that the clamp loader of the Escherichia coli replicase has the composition DnaX 3 ’. Two DnaX proteins exist in E. coli , full length and a truncated that is created by ribosomal frameshifting. binds DNA polymerase III tightly; does not. There is a controversy as to whether or not DNA polymerase III holoenzyme (Pol III HE) contains . A three- form of Pol III HE would contain three Pol IIIs. Proponents of the three- hypothesis have claimed that found in Pol III HE might be a proteolysis product of . To resolve this controversy, we constructed a strain that expressed only from a mutated chromosomal dnaX . containing a C-terminal biotinylation tag (-C tag ) was provided in trans at physiological levels from a plasmid. A 2000-fold purification of Pol III* (all Pol III HE subunits except β) from this strain contained one molecule of -C tag per Pol III* assembly, indicating that the dominant form of Pol III* in cells is Pol III 2 2 ’. Revealing a role for in cells, mutants that express only display sensitivity to ultraviolet light and reduction in DNA Pol IV-dependent mutagenesis associated with double-strand-break repair, and impaired maintenance of an F’ episome.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
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