PubMed HealthSearch

Biomedical subjects

S Kalyanaraman

Publications and source records attributed to S Kalyanaraman.

At least 19 recordsLinked to original sources

A brain-specific G protein gamma subunit.

Two different cDNAs for G protein gamma subunits have been isolated from mouse brain. One encodes a novel gamma subunit, gamma 4, the expression of which is detected only in brain. A fragment of this cDNA had been isolated previously. The other cDNA encodes gamma 3, a subunit type previously isolated from bovine brain. The primary structure of the gamma 3 subunit is conserved completely across species indicating that the diversity in the structure of the gamma subunits is of functional consequence. Moreover, gamma 2, gamma 3 and gamma 4, which are predominantly expressed in brain, are more homologous to each other than other gamma subunits, indicating that the G protein gamma subunits belong to distinct subfamilies similar to the alpha subunits.

Amino Acid Sequence

Generation of hybrid human immunodeficiency virus by homologous recombination.

Human immunodeficiency virus (HIV) type 1, isolated from diverse sources, exhibits genomic diversity. The mechanisms by which the genomic diversity takes place in individuals exposed to multiple virus isolates is yet to be elucidated. Genetic variation, in general, might result from mutagenic events such as point mutations, rearrangements (insertions and deletions), and recombination. In an attempt to evaluate the process of genetic diversity, we designed experiments to analyze recombination between HIV DNAs by using DNA transfection in cell cultures. Here we report the successful recombination between truncated HIV proviral DNAs with an overlap homology of 53 base pairs that leads to the formation of viable hybrid virus. Recombination was also seen between exogenous DNA introduced into cells and homologous HIV sequences resident in the cells. These results indicate that recombination among various HIV isolates may play a significant role in the generation of genetic diversity of HIV. Further, the method used here enables the construction of hybrid HIV genomes to identify the viral determinants responsible for tropism, replication, and cytopathic effects.

Base Sequence

Homologous recombination between human immunodeficiency viral DNAs in cultured human cells: analysis of the factors influencing recombination.

Recombination between HIV DNAs was analyzed using DNA transfection in cell cultures and the optimal conditions for efficient recombination were determined. Recombinant plasmid DNA substrates were constructed from HIV proviral DNAs and the success of recombination was measured by the production of viable hybrid virus. The process of recombination between HIV DNAs was shown to be i) dependent on homology between the truncated HIV DNAs and ii) maximum with concentrations of the truncated DNAs 3ug and above. HIV isolates with heterogeneity in their primary sequence, thus offer an ideal system for the analysis of the requirement of homologous recombination. In addition, recombination methodology would be useful for generating hybrid HIVs for the analysis of specific viral gene functions.

Calcium Phosphates

Double-stranded RNA specific nuclease from germinating embryos of Pennisetum typhoides.

A double-stranded RNA specific nuclease (ds RNase) has been purified from the pearl millet Pennisetum typhoides. The purification involved S-30 preparation from the germinating embryos, DEAE-cellulose and DNA-cellulose chromatography. The partially pure enzyme preferentially solubilized the synthetic double-stranded polynucleotide [3H]poly(rA) . poly(rU); the degradation of [3H]poly(rC) was fourteen fold lower under the same assay conditions. Furthermore, the ds RNase activity was inhibited to an extent of 58% by ethidium bromide, which is known to intercalate with double-stranded RNAs. Active sulfhydryl groups were found to be necessary for the ds RNase activity since the enzyme action was inhibited by N-ethylmaleimide. Ethidium bromide and N-ethyl-maleimide did not significantly inhibit the ss RNase activity. In contrast, diethyl pyrocarbonate inhibited ss RNase activity completely and ds RNase by 58%. Heating the enzyme for 20 min at 50 degrees C resulted in drastic loss of both enzyme activities. The ds RNase showed maximum activity in the pH range of 6.5 to 7.5. The enzyme acts in vitro on E. coli 30S precursor ribosomal RNA and the cleavage products migrated in the region of mature 23S and 16S rRNAs.

Endoribonucleases

Partial purification and characterization of a double-stranded RNA-specific nuclease from human placenta.

A double-stranded RNA-specific nuclease (ds RNase) has been isolated and partially purified from human placenta by DEAE-cellulose and DNA-cellulose column chromatography. Denatured DNA-cellulose retained most of the single-stranded RNA-specific nuclease (ss RNase) activity, whereas the ds RNase came out in the void volume. N-ethylmaleimide at a concentration of 5 mM, selectively inhibited ds RNase activity by 60% under the conditions in which the ss RNase activity was inhibited to an extent of 7%. The ds RNase was specifically inhibited by Penicillium chrysogenum viral ds RNA and by ethidium bromide. The partially purified ds RNase showed requirements for Mg+ whereas Mn2+ and NH4+ ions were inhibitory. The DEAE-enzyme cleaved 32P-labelled 45S ribosomal precursor RNAs from Yoshida ascites sarcoma cells into species that had similar electrophoretic mobilities as the mature rRNAs.

Ammonia