Thermodynamic, Spectroscopic, and Equilibrium Binding Studies of DNA Sequence Context Effects in Six 22-Base Pair Deoxyoligonucleotides

Biochemistry ◽  
1999 ◽  
Vol 38 (34) ◽  
pp. 11197-11208 ◽  
Author(s):  
Peter V. Riccelli ◽  
Peter M. Vallone ◽  
Irina Kashin ◽  
Brian D. Faldasz ◽  
Michael J. Lane ◽  
...  
Keyword(s):  
Dna Sequence ◽  
Base Pair ◽  
Context Effects ◽  
Binding Studies ◽  
Sequence Context ◽  
Equilibrium Binding

scholarly journals DNA Sequence Context Effects on the Glycosylase Activity of Human 8-Oxoguanine DNA Glycosylase

2012 ◽  
Vol 287 (44) ◽  
pp. 36702-36710 ◽  
Author(s):  
Akira Sassa ◽  
William A. Beard ◽  
Rajendra Prasad ◽  
Samuel H. Wilson
Keyword(s):  
Dna Sequence ◽  
Context Effects ◽  
Dna Glycosylase ◽  
Sequence Context

The yeast rad18 mutator specifically increases G.C----T.A transversions without reducing correction of G-A or C-T mismatches to G.C pairs

1991 ◽  
Vol 11 (1) ◽  
pp. 218-225
Author(s):  
B A Kunz ◽  
X L Kang ◽  
L Kohalmi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Dna Sequence ◽  
Base Pair ◽  
Mutation Frequency ◽  
Wild Type ◽  
Sequence Context ◽  
Single Base ◽  
Mutator Effect ◽  
Single Base Pair

Inactivation of the Saccharomyces cerevisiae RAD18 gene confers a mutator phenotype. To determine the specificity of this effect, a collection of 212 spontaneous SUP4-o mutants arising in a rad18 strain was characterized by DNA sequencing. Comparison of the resulting mutational spectrum with that for an isogenic wild-type (RAD18) strain revealed that the rad18 mutator specifically enhanced the frequency of single base pair substitutions. Further analysis indicated that an increase in the frequency of G.C----T.A transversions accounted for the elevated SUP4-o mutation frequency. Thus, rad18 is the first eucaryotic mutator found to generate only a particular base pair substitution. The majority of G.C pairs that were not mutated in the rad18 background were at sites where G.C----T.A events can be detected in SUP4-o, suggesting that DNA sequence context influences the rad18 mutator effect. Transformation of heteroduplex plasmid DNAs into the two strains demonstrated that the rad18 mutator did not reduce the efficiency of correcting G-A or C-T mismatches to G.C pairs or preferentially correct the mismatches to A.T pairs. We propose that the RAD18 gene product might contribute to the fidelity of DNA replication in S. cerevisiae by involvement in a process that serves to limit the formation of G-A and C-T mismatches at template guanine and cytosine sites during DNA synthesis.

scholarly journals The yeast rad18 mutator specifically increases G.C----T.A transversions without reducing correction of G-A or C-T mismatches to G.C pairs.

1991 ◽  
Vol 11 (1) ◽  
pp. 218-225 ◽  
Author(s):  
B A Kunz ◽  
X L Kang ◽  
L Kohalmi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Dna Sequence ◽  
Base Pair ◽  
Mutation Frequency ◽  
Wild Type ◽  
Sequence Context ◽  
Single Base ◽  
Mutator Effect ◽  
Single Base Pair

Inactivation of the Saccharomyces cerevisiae RAD18 gene confers a mutator phenotype. To determine the specificity of this effect, a collection of 212 spontaneous SUP4-o mutants arising in a rad18 strain was characterized by DNA sequencing. Comparison of the resulting mutational spectrum with that for an isogenic wild-type (RAD18) strain revealed that the rad18 mutator specifically enhanced the frequency of single base pair substitutions. Further analysis indicated that an increase in the frequency of G.C----T.A transversions accounted for the elevated SUP4-o mutation frequency. Thus, rad18 is the first eucaryotic mutator found to generate only a particular base pair substitution. The majority of G.C pairs that were not mutated in the rad18 background were at sites where G.C----T.A events can be detected in SUP4-o, suggesting that DNA sequence context influences the rad18 mutator effect. Transformation of heteroduplex plasmid DNAs into the two strains demonstrated that the rad18 mutator did not reduce the efficiency of correcting G-A or C-T mismatches to G.C pairs or preferentially correct the mismatches to A.T pairs. We propose that the RAD18 gene product might contribute to the fidelity of DNA replication in S. cerevisiae by involvement in a process that serves to limit the formation of G-A and C-T mismatches at template guanine and cytosine sites during DNA synthesis.

scholarly journals Kinetic mechanism of Na+-coupled aspartate transport catalyzed by GltTk

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Gianluca Trinco ◽  
Valentina Arkhipova ◽  
Alisa A. Garaeva ◽  
Cedric A. J. Hutter ◽  
Markus A. Seeger ◽  
...  
Keyword(s):  
Kinetic Mechanism ◽  
Protein Quantification ◽  
Detergent Solution ◽  
Sodium Ions ◽  
Binding Studies ◽  
Equilibrium Binding ◽  
Uptake Rates ◽  
Bona Fide ◽  
Inside Out

AbstractIt is well-established that the secondary active transporters GltTk and GltPh catalyze coupled uptake of aspartate and three sodium ions, but insight in the kinetic mechanism of transport is fragmentary. Here, we systematically measured aspartate uptake rates in proteoliposomes containing purified GltTk, and derived the rate equation for a mechanism in which two sodium ions bind before and another after aspartate. Re-analysis of existing data on GltPh using this equation allowed for determination of the turnover number (0.14 s−1), without the need for error-prone protein quantification. To overcome the complication that purified transporters may adopt right-side-out or inside-out membrane orientations upon reconstitution, thereby confounding the kinetic analysis, we employed a rapid method using synthetic nanobodies to inactivate one population. Oppositely oriented GltTk proteins showed the same transport kinetics, consistent with the use of an identical gating element on both sides of the membrane. Our work underlines the value of bona fide transport experiments to reveal mechanistic features of Na+-aspartate symport that cannot be observed in detergent solution. Combined with previous pre-equilibrium binding studies, a full kinetic mechanism of structurally characterized aspartate transporters of the SLC1A family is now emerging.

scholarly journals DNA Sequence Context as a Determinant of the Quantity and Chemistry of Guanine Oxidation Produced by Hydroxyl Radicals and One-electron Oxidants

2008 ◽  
Vol 283 (51) ◽  
pp. 35569-35578 ◽  
Author(s):  
Yelena Margolin ◽  
Vladimir Shafirovich ◽  
Nicholas E. Geacintov ◽  
Michael S. DeMott ◽  
Peter C. Dedon
Keyword(s):  
Dna Sequence ◽  
Hydroxyl Radicals ◽  
Sequence Context ◽  
Guanine Oxidation

Sequence Context Effects on Mutational Properties of cis-Opened Benzo[c]phenanthrene Diol Epoxide−Deoxyadenosine Adducts in Site-Specific Mutation Studies†

Biochemistry ◽  
1999 ◽  
Vol 38 (3) ◽  
pp. 1144-1152 ◽  
Author(s):  
Ingrid Pontén ◽  
Jane M. Sayer ◽  
Anthony S. Pilcher ◽  
Haruhiko Yagi ◽  
Subodh Kumar ◽  
...  
Keyword(s):  
Context Effects ◽  
Sequence Context ◽  
Site Specific ◽  
Specific Mutation ◽  
Diol Epoxide ◽  
Deoxyadenosine Adducts

DNA sequence analysis of spontaneous mutations in the SUP4-o gene of Saccharomyces cerevisiae

1988 ◽  
Vol 8 (2) ◽  
pp. 978-981
Author(s):  
C N Giroux ◽  
J R Mis ◽  
M K Pierce ◽  
S E Kohalmi ◽  
B A Kunz
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sequence Analysis ◽  
Transposable Elements ◽  
Dna Sequencing ◽  
Dna Sequence ◽  
Base Pair ◽  
Dna Sequence Analysis ◽  
Spontaneous Mutations ◽  
Yeast Saccharomyces Cerevisiae ◽  
Spontaneous Mutagenesis

A collection of 196 spontaneous mutations in the SUP4-o gene of the yeast Saccharomyces cerevisiae was analyzed by DNA sequencing. The classes of mutation identified included all possible types of base-pair substitution, deletions of various lengths, complex alterations involving multiple changes, and insertions of transposable elements. Our findings demonstrate that at least several different mechanisms are responsible for spontaneous mutagenesis in S. cerevisiae.

Repetitive Dictyostelium heat-shock promotor functions in Saccharomyces cerevisiae

1984 ◽  
Vol 4 (4) ◽  
pp. 591-598
Author(s):  
J Cappello ◽  
C Zuker ◽  
H F Lodish
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Nucleotide Sequence ◽  
Dna Sequence ◽  
Base Pair ◽  
Shock Treatment ◽  
Yeast Cells ◽  
Genomic Segment ◽  
Heat Shock Treatment ◽  
Shock Response

The Dictyostelium genome contains 40 copies of a 4.7-kilobase repetitive and apparently transposable DNA sequence (DIRS-1) and about 250 smaller elements that appear to be deletions or rearrangements of DIRS-1. Transcripts of these sequences are induced during differentiation and also by heat shock treatment of growing cells. We showed that one such cloned element, pB41.6 (2.5 kilobases) contains a nucleotide sequence identical to the Drosophila consensus heat shock promotor. To test whether this sequence might indeed control the expression of DIRS-1-related RNAs, we have cloned this genomic segment into yeast cells. In yeast cells, 41.6 directs synthesis of a 1.7-kilobase RNA that is induced at least 10-fold by heat shock. Transcription initiates at about 124 bases 3' of the putative promotor sequence and terminates within the 41.6 insert. A 381-base-pair subclone that contains the putative promotor sequence is sufficient to induce the heat shock response of 41.6 in yeast cells.

scholarly journals A plant DNA-binding protein that recognizes 5-methylcytosine residues.

1989 ◽  
Vol 9 (3) ◽  
pp. 1351-1356 ◽  
Author(s):  
D L Zhang ◽  
K C Ehrlich ◽  
P C Supakar ◽  
M Ehrlich
Keyword(s):  
Dna Binding ◽  
Dna Sequence ◽  
Base Pair ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Sequence Specificity ◽  
Dna Sequence Specificity ◽  
Plant Dna ◽  
Related Family ◽  
Nuclear Extracts

A novel, 5-methylcytosine-specific, DNA-binding protein, DBP-m, has been identified in nuclear extracts of peas. DBP-m specifically recognizes 5-methylcytosine residues in DNA without appreciable DNA sequence specificity, unlike a mammalian DNA-binding protein (MDBP), which recognizes 5-methylcytosine residues but only in a related family of 14-base-pair sequences.

Sign in / Sign up

Export Citation Format

Share Document