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The nucleotide sequence of a rat 18 S ribosomal ribonucleic acid gene and a proposal for the secondary structure of 18 S ribosomal ribonucleic acid.

The nucleotide sequence of a rat 18 S rRNA gene was determined. The 18 S rRNA encoded in the gene contains 1874 nucleotides, and the molecular weight estimated from the sequence is 6.09 X 10(5). The sequences of rat and Xenopus laevis 18 S rRNAs are very similar; the only differences of consequence are the insertions between nucleotides 197 and 206 and between 258 and 279 of the rat nucleic acid of sequences rich in guanine and cytosine. A proposal is presented for the secondary structure of rat 18 S rRNA based on a comparison with the sequences of 17 other small ribosomal subunit RNAs. While the primary sequences of rat and eubacterial RNAs are different, the deduced secondary structures are remarkably similar.

Animals↗

Reinitiation of deoxyribonucleic acid synthesis by deoxyribonucleic acid initiation mutants of Escherichia coli: role of ribonucleic acid synthesis, protein synthesis, and cell division.

The dnaA and dnaC genes are thought to code for two proteins required for the initiation of chromosomal deoxyribonucleic acid replication in Escherichia coli. When a strain carrying a mutation in either of these genes is shifted from a permissive to a restrictive temperature, chromosome replication ceases after a period of residual synthesis. When the strains are reincubated at the permissive temperature, replication again resumes after a short lag. This reinitiation does not require either protein synthesis (as measured by resistance to chloramphenicol) or ribonucleic acid synthesis (as measured by resistance to rifampin). Thus, if there is a requirement for the synthesis of a specific ribonucleic acid to initiate deoxyribonucleic acid replication, this ribonucleic acid can be synthesized prior to the time of initiation and is relatively stable. Furthermore, the synthesis of this hypothetical ribonucleic acid does not require either the dnaA of dnaC gene products. The buildup at the restrictive temperature of the potential to reinitiate deoxyribonucleic acid synthesis at the permissive temperature shows rather complex kinetics the buildup roughly parallels the rate of mass increase of the culture for at least the first mass doubling at the restrictive temperature. At later times there appears to be a gradual loss of initiation potential despite a continued increase in mass. Under optimal conditions the increase in initiation potential can equal, but not exceed, the increase in cell division at the restrictive temperature. These results are most easily interpreted according to models that postulate a relationship between the initiation of deoxyribonucleic acid synthesis and the processes leading to cell division.

Bacterial Proteins↗

Acquisition of native conformation of ribosomal 5S ribonucleic acid from Escherichia coli. Hydrodynamic and spectroscopic studies on the unfolding and refolding of ribonucleic acid.

In a continuing effort to decipher the molecular mechanism of ribosome self-assembly [e.g., Dunn, J. M., & Wong, K.-P. (1979) Biochemistry 18, 4380-4385], the mechanism of folding of 5S RNA was investigated by unfolding and refolding studies using several physical techniques including circular dichroism (CD), UV absorption spectroscopy, and sedimentation velocity analysis to monitor various conformational changes. The 5S RNA was unfolded by using 6 M urea and EDTA, and an unfolded state was characterized in which the base pairing was found to be disrupted, but extensive base stacking remained. The unfolded 5S RNA was then refolded upon removal of urea and EDTA by dialysis against a reconstitution buffer both with and without Mg2+, and the refolded states were characterized. The results indicate that under the proper conditions, 5S RNA refolds to a conformation and overall shape very similar to the native conformation. These results indicate that the nucleotide sequence in 5S RNA contains the necessary information to direct the folding of the RNA into its native conformation. The presence of an appropriate concentration of Mg2+ and an incubation at 60 degrees C are required for the correct refolding, since omission of either one results in a renatured 5S RNA whose conformation is quite different from the native one.

Circular Dichroism↗

Regulation of ribonucleic acid synthesis by polyamines. Reversal by spermine of inhibition by methylglyoxal bis(guanylhydrazone) of ribonucleic acid synthesis and histone acetylation in rabbit heart.

The relationship between polyamines and RNA synthesis was studied by considering the action of spermine on histone acetylation in perfused heart. In addition, the effect of methylglyoxal bis(guanylhydrazone), inhibitor of putrescine-activated S-adenosylmethionine decarboxylase activity, on RNA and polyamine specific radioactivity and on acetylation of histone fractions was also investigated in perfused heart. Different concentrations of spermine and/or methylglyoxas bis(guanylhydrazone) were injected into the heart, 15 min after beginning the perfusion. The results demonstrate that spermine stimulates the specific radioactivity of RNA of subcellular fractions. Acetylation of the arginine-rich histone fractions, involved in the regulation of RNA transcription, is enhanced by spermine. The perfusion with methylglyoxal bis(guanylhydrazone) causes a decrease in the specific radioactivity of polyamines and RNA, and in acetylation of histone fractions. However, spermine is able to reverse the methylglyoxal bis(guanylhydrazone) inhibition when injected simultaneously. From these results we may assume a possible role for spermine in the regulation of RNA transcription.

Acetylation↗

Replication of bacteriophage ribonucleic acid: some physical properties of single-stranded, double-stranded, and branched viral ribonucleic acid.

Replicative intermediate (RI) is considered to be the double-stranded ribo-nucleic acid (RNA) template for synthesis of viral RNA, with bound nascent single-stranded viral RNA. A theoretical description of RI is based on the analysis of a steady state of biopolymerization on a template which determines not only nucleotide sequence, but also chain length. The hydrodynamic properties of RI isolated from Escherichia coli infected with bacteriophage R17 are compared with those of RNA isolated from R17 (single-stranded RNA) and of replicative form (RF) isolated from E. coli infected with R17. RF is double-stranded RNA template without any single-stranded component. Whereas S for R17 RNA is a function of the ionic strength (Gamma/2) of the solvent, S is almost invariant with Gamma/2 for RF. By contrast S for RI lies between the sedimentation constants for R17 RNA and RF and S varies with Gamma/2 as does R17 RNA. The weight distribution of S for RI demonstrates the heterogeneity of this material, and the variation in the weight distribution with ionic strength demonstrates the duality of structure in RI. Using S and [eta], the Mw for RI is estimated to be 2.6 x 10(6) daltons, as compared with the theoretical value of 2.9 x 10(6) daltons.

Coliphages↗