Chapter 19 DNA-Binding Proteins

Stein, Gretchen H.

doi:10.1016/s0091-679x(08)61148-6

Cited by 7 publications

(3 citation statements)

References 40 publications

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“…However, a number of other types of support can be used and may have dramatic effects on purifications obtained [for reviews, see Allfrey & Inoue (1978) and Stein (1978)]. Allfrey et al have prepared DNA-Sephadex columns using carbo-diimide under conditions which favor the formation of linkages between dextran hydroxyls and monoesterified phosphates [Allfrey et al, 1973; see also Gilham (1968) andWeissbach &Poonian (1974)].…”

Section: Discussionmentioning

confidence: 99%

Isolation of a protein fraction that binds preferentially to chicken middle repetitive DNA

Sanzo¹,

Stevens²,

Tsai³

et al. 1984

Biochemistry

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We have fractionated oviduct tissue extracts by using a combination of ion-exchange and DNA-Sephadex chromatography. By comparing the electrophoretic patterns of proteins eluted from competing specific and nonspecific DNA columns, we isolated a fraction which bound with specificity to columns containing the chicken middle repetitive sequence "CR1". This fraction showed a clear preference for binding to separate, cloned CR1 fragments derived from either the 5' or the 3' transition region of the ovalbumin gene domain when examined by using nitrocellulose filter binding assays. To localize the protein binding site, a CR1 clone was digested with various restriction enzymes, and the resulting fragments were examined for preferential protein binding. Results suggest that the binding site lies within a 39-nucleotide sequence which is highly conserved among different CR1 elements. This finding represents the first isolation of a protein which demonstrates a preference for binding to a middle repetitive sequence and suggests that this interaction may have a biological role. The DNA column competition adsorption method should have general application to the isolation of other gene-regulating proteins possessing DNA sequence preference.

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Section: Discussionmentioning

confidence: 99%

Isolation of a protein fraction that binds preferentially to chicken middle repetitive DNA

Sanzo¹,

Stevens²,

Tsai³

et al. 1984

Biochemistry

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“…Double-stranded DNA-cellulose was prepared from Sigma type V calf thymus DNA and Bio-Rad Cellex 410 as described (Alberts & Herrick, 1971) with the addition of a UV irradiation step (Litman, 1968). The chromatographic matrix contained 0.42 mg of DNA/mL of packed cellulose as determined by digestion with DNAse I and hydrolysis with 0.5 M perchloric acid (Stein, 1978). For chromatography, the method of Alberts & Herrick (1971) was adapted for gradient elution of HI.…”

Section: Removal Of Noncovalently Bound [}H]b[a]p and Othermentioning

confidence: 99%

Binding of benzo[a]pyrene to histones and altered affinity of modified histone 1 for DNA

Jenson¹,

Fitzgibbon²,

Brennan³

et al. 1982

Biochemistry

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The covalent binding of benzo[a]pyrene (B[a]P) to acid extractable chromosomal proteins and the subsequent effect on histone 1-DNA interaction have been characterized in a model system by utilizing calf thymus nuclei as targets and rat liver microsomes as an exogenous source of enzymes for the metabolic activation of B[a]P. A two-step ion-exchange chromatography and desalting procedure was employed for removing noncovalently bound B[a]P and other contaminants. Fluorography of acetic acid-urea and Triton-acetic acid-urea-polyacrylamide gels indicated that H1 and H3 were the only principal histone targets in [3H]B[a]P-modified calf thymus nuclei. The validity of this assignment was confirmed by comparison of the chromatographic distributions of [3H]B[a]P cpm among peptides derived from the HClO4- soluble (H1) and HClO4-insoluble (core histones) protein fractions to the distributions obtained for authentic individual histone fractions. Comparison of amino acid compositions in individual peptide fractions which bound [3H]B[a]P differentially yielded some insight into the probable target amino acid residues for B[a]P binding. On the basis of electrophoresis in polyacrylamide gels, it appeared as if B[a]P had bound to multiple subfractions of H1 and H3. The equivalent distribution of covalently attached [3H]B[a]P among the major peptides of H1 and H3 modified either in intact nuclei or while free in solution implied that the relative accessibility of major portions of the H1 and H3 molecules for covalent B[a]P binding is not affected by interactions with DNA or other chromosomal proteins. Covalent attachment of [3H]B[a]P to purified H1 reduced the affinity of this histone for DNA-cellulose.

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“…The eukaryotic nucleus is associated with some 250-500 proteins visible by Coomassie blue staining of acrylamide gels (1,2). These visible proteins are certainly present in many copies per cell.…”

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confidence: 99%

Tissue-specific and species-specific monoclonal antibodies to avian red cell nuclear proteins.

Kane¹,

Cheng²,

Burch³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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Chapter 19 DNA-Binding Proteins

Cited by 7 publications

References 40 publications

Isolation of a protein fraction that binds preferentially to chicken middle repetitive DNA

Isolation of a protein fraction that binds preferentially to chicken middle repetitive DNA

Binding of benzo[a]pyrene to histones and altered affinity of modified histone 1 for DNA

Tissue-specific and species-specific monoclonal antibodies to avian red cell nuclear proteins.

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