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R J Mans

Publications and source records attributed to R J Mans.

11 recordsLinked to original sources

Conservation of the primary structure at the 3' end of 18S rRNA from eucaryotic cells.

DNA sequencing methods have been used to determine a sequence of about 20 nucleotides at the 3' termini of various 18S (small ribosomal subunit) RNA molecules. Polyadenylated rRNA was first synthesized using the enzyme ATP:polynucleotidyl transferase from mainze. Then in the presence of an oligonucleotide primer uniquely complementary to the end of each adenylated rRNA, a cDNA copy was produced using AMV reverse transcriptase. In every case, the cDNA transcript was of finite size, which we ascribe to the appearance of an oligonucleotide containing m62A near the 3' end of the 18S rRNAs. Sequences at the 3' termini of 18S rRNA molecules from the four eucaryotic species examined here (mouse, silk worm, wheat embryo and slime mold) are highly conserved. They also exhibit strong homology to the 3' end of E. coli 16S rRNA. Two important differences, however, are apparent. First, the 16S sequence CCUCC, implicated in mRNA binding by E. coli ribosomes, is absent from each eucaryotic rRNA sequence. Second, a purine-rich region which exhibits extensive complementarity to the 5' noncoding regions of many eucaryotic mRNAs appears consistently.

Animals

Utilization of ribonucleic acid and deoxyoligomer primers for polyadenylic acid synthesis by adenosine triphosphate: polynucleotidylexotransferase from maize.

The ATP:polynucleotidylexotransferase isolated and purified from maize seedlings catalyzes the synthesis of polyadenylic acid by the sequential addition of 80 to 200 AMP moieties from ATP to the 3'-hydroxyl terminus of either ribo- or deoxyoligomers. Copurification of the RNA and DNA-primed activities, identical metal cofactor and reaction requirements for either primer and identical heat inactivation curves with either primer strongly suggest that both primers are utilized by the same enzyme.

Adenosine Triphosphate

Evidence for fidelity of chromatin reconstitution.

Several lines of evidence are presented which support the contention that chromatin may be dissociated, fractionated, and reconstituted without altering the compositional, structural, or transcriptional integrity of the genome. The similar compositions of native and reconstituted chromatins are suggested by the absence of significant differences in their protein/DNA ratios and in the polyacrylamide gel electrophoretic profiles of their histones and nonhistone chromosomal proteins. Criteria for fidelity of genome structure in reconstituted chromatin include binding of reporter molecules with specificity for the minor groove of DNA, binding of histones, number of sites available for addition of nucleotides, and circular dichroism spectra. When the transcriptional activities of native and reconstituted chromatins were compared under conditions where reinitiation is prohibited, significant changes were not observed. Taken together, the present results strongly suggest, but do not conclusively establish, fidelity of chromatin reconstitution.

Animals