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W E Timberlake

Publications and source records attributed to W E Timberlake.

85 records · Page 5Linked to original sources
PubMed1978

Sequence complexity of nuclear and polysomal RNA in leaves of the tobacco plant.

The first measurements are reported of the sequence complexity of nuclear and polysomal RNA contained within the cells of a higher plant. Polysomal RNA from tobacco leaves was prepared by a procedure which minimized contamination with nuclear RNA. Hybridization of 3H-cDNA complementary to polysomal poly(A) RNA with an excess of tobacco DNA indicated that greater than 95% of the poly(A) mRNA was transcribed from single-copy sequences. RNA excess hybridization reactions with polysomal poly(A) RNA and 3H-cDNA revealed the presence of three abundance classes in the poly(A) mRNA. The best least-squares solution indicated that these classes comprise 9, 52, and 39% of the poly (A) mRNA and contain sequences present an average of 4500, 340 and 17 times per cell. Hybridization reactions containing an excess of nuclear or total polysomal RNA and 3H-single-copy DNA indicated the complexity of these RNA populations to be 1.19 X 10(8) nucleotides (nuclear) and 3.33 X 10(7) nucleotides (polysomal). Thus only 28% of the nuclear RNA sequence diversity (27,000 average-sized mRNA sequences) is respresented in leaf polysomes. These results suggest that there is a general similarity in the basic transcriptional processes of metaphytan and metazoan cells.

Base Sequence↗
PubMed1977

Relationship between nuclear and polysomal RNA populations of Achlya: a simple eucaryotic system.

The relationship between hnRNA and mRNA in the water mold Achlya has been investigated in several ways. Analysis of the nuclear and polysomal poly(A) RNA by sucrose density gradient centrifugation under denaturing and nondenaturing conditions showed that the populations have indistinguishable size distributions. The number average sizes were calculated to be 1150 nucleotides for nuclear and 1140 nucleotides for polysomal poly(A) RNA. Selective inhibition of rRNA synthesis was used to investigate the size distribution of hnRNA without regard to poly(A) content. Very little hnRNA was observed which sedimented more rapidly than polysomal poly(A) RNA. Hybridization experiments in which an excess of nuclear DNA was reacted with 3H-poly(A) hnRNA or 3H-poly(A) mRNA showed that both populations contain repetitive transcripts (9-10%) as well as single-copy transcripts (44%). Analysis of hybrids on hydroxyapatite in the presence of 8 M urea demonstrated that the poly(A) RNA complementary to repetitive DNA sequence components represented a population of molecules distinct from the population complementary to single-copy DNA. The complexity of whole cell, nuclear and polysomal RNA was determined by saturation hypbridization to single-copy 3H-DNA. All three populations were complementary to essentially the same fraction of the DNA. Terminal hybridization values were 3.84, 3.76 and 3.76% for whole cell, nuclear and polysomal RNA, respectively, representing a complexity of 2.1 X 10(6) nucleotides. These data suggest that the composition of the hnRNA and mRNA populations are essentially identical. No evidence for selective turnover of any sequence component or size class within the nucleus was observed.

Base Sequence↗
PubMed1977

DNA sequence organization in the water mold Achlya.

Experiments are described that characterize the organization of DNA sequences in the water mold Achlya bisexualis. These experiments demonstrate that repetitive and single copy sequences in the Achlya genome are arranged in a long-period interspersion pattern. Estimates of the spacing intervals between repetitive and single copy DNA indicate, however, that the interspersion pattern in Achlya is longer than has been previously reported in other eukaryotes. These data and measurements of structural gene expression in Achlya [Timberlake, W.E., Shumard, D. S. & Goldberg, R. B. (1977) Cell 10, 623-632] make it difficult to propose a regulatory function for repeated DNA in this eukaryote.

Journal Article↗
PubMed1974

Differential effects of analogs of cycloheximide on protein and RNA synthesis in Achlya.

Analogs of the glutarimide antibiotic cycloheximide were tested for their effect on growth and incorporation of proline and uridine into acid-insoluble material in Achlya bisexualis. Each of the compounds tested had reduced antibiotic activity as compared to cycloheximide. The effects of the antibiotics on protein and RNA synthesis were varied. While cycloheximide inhibited both protein and RNA synthesis immediately, two of the analogs inhibited proline incorporation without effect on uridine incorporation, while three, each representing a modification of the hydroxyl of cycloheximide, stimulated uridine incorporation and either had no effect on or inhibited protein synthesis. These results indicate that the control of RNA synthesis by protein synthesis in Achlya can be released by glutarimide antibiotics.

Antifungal Agents↗
PubMed1973

Protein synthesis during the differentiation of sporangia in the water mold Achlya.

During the synchronous differentiation of sporangia in the absence of added nutrients, the water mold Achlya bisexualis (Coker and Couch) actively synthesized protein. Inhibition of protein synthesis at any time during the sporulation process completely inhibited further differentiation. Large changes in the rate of radioactive amino acid uptake resulted in changes in the specific activity of the cellular amino acid pool. The rate of protein synthesis was calculated from the amino acid pool specific activity and the incorporation of isotope into protein. During the 1st h after induction of the sporulation process, the rate of protein synthesis increased to two times the initial value. The amino acid precursors for this synthesis were supplied by the degradation of preexisting protein. Proteolytic enzyme activity assayed in vitro increased in proportion to the in vivo rates of protein synthesis and degradation. Differentiation was accompanied by a slight decline in dry weight of the mycelium as well as by a decrease in the protein content, whereas the relative size of the amino acid pools remained constant.

Amino Acids↗