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J Tower

Publications and source records attributed to J Tower.

27 records · Page 2Linked to original sources
PubMed1987

Transcription of mouse rDNA is regulated by an activated subform of RNA polymerase I.

We have identified the species-nonspecific factor required for mouse rDNA transcription, factor C, as an activated subform of RNA polymerase I. C is an RNA polymerase I since it copurifies with bulk polymerase I activity on the three chromatographic columns used to achieve a virtually homogenous preparation of polymerase I, as well as on four additional matrices; it is quantitatively neutralized as well as immunoprecipitated by two different types of anti-polymerase I antibodies; and it has thermal lability identical to that of bulk polymerase I. However, C is clearly distinct from bulk polymerase I in its ability to participate in the stable rDNA transcription complex and to catalyze accurate initiation of rRNA synthesis. It also has a greater sedimentation coefficient than bulk polymerase I. Furthermore, this activated polymerase subform is specifically lacking in extracts of cells in which rDNA transcription was down-regulated because of cycloheximide treatment or attainment of stationary phase. These data suggest that regulation of rDNA transcription in vivo may involve modulation in availability of the activated polymerase I subform.

Animals↗
PubMed1986

Factors and nucleotide sequences that direct ribosomal DNA transcription and their relationship to the stable transcription complex.

We have studied the protein components and nucleic acid sequences involved in stably activating the ribosomal DNA (rDNA) template and in directing accurate transcription of mammalian rRNA genes. Two protein components are necessary to catalyze rDNA transcription, and these have been extensively purified. The first, factor D, can stably associate by itself with the rDNA promoter region and is responsible for template commitment. The second component, factor C, which appears to be an activated subset of polymerase I, can stably bind to the factor D-rDNA complex but not to the rDNA in the absence of factor D. A third component which had been previously identified as a rDNA transcription factor is shown to be a RNase inhibitor. Extending our earlier observation that the approximately 150-base-pair mouse rDNA promoter consists of a minimal essential region (residues approximately -35 to approximately +9) and additional upstream stimulatory domains, we now report that each of these promoter domains acts to augment the binding of the polymerase I transcription factors. A minimum core region (residues approximately -35 to approximately -15) is capable of stable complex formation and of binding transcription factor D. Factor C can also bind to this D-core region complex.

Animals↗
PubMed1985

A complex control region of the mouse rRNA gene directs accurate initiation by RNA polymerase I.

To determine the size and location of the mouse rDNA promoter, we constructed systematic series of deletion mutants approaching the initiation site from the 5' and 3' directions. These templates were transcribed in vitro under various conditions with S-100 and whole-cell extracts. Surprisingly, the size of the rDNA region that determines the level of transcription differed markedly, depending on the reaction conditions. In both kinds of cell extracts, the apparent 5' border of the promoter was at residue ca. -27 under optimal transcription conditions, but as reaction conditions became less favorable, the 5' border moved progressively out to residues -35, -39, and -45. The complete promoter, however, extends considerably further, for under other nonoptimal conditions, we observed major effects of promoter domains extending in the 5' direction to positions ca. -100 and -140. In contrast, the apparent 3' border of the mouse rDNA promoter was at residue ca. +9 under all conditions examined. We also show that the subcloned rDNA region from -39 to +9 contains sufficient information to initiate accurately and that the region between +2 and +9 can influence the specificity of initiation. These data indicate that, although the polymerase I transcription factors recognize and accurately initiate with only the sequences downstream of residue -40, sequences extending out to residue -140 greatly favor the initiation reaction; presumably, this entire region is involved in rRNA transcription in vivo.

Animals↗
PubMedDate not supplied

Sequence requirements for upregulated expression of Drosophila hsp70 transgenes during aging.

hsp70 protein and hsp70:lacZ fusion reporters are upregulated during aging and in response to oxidative stress in the thorax of Drosophila. hsp70 expression was increased during aging in each of seven different Drosophila genetic backgrounds tested, 2.6-4.8-fold. DNA sequence requirements were investigated by analysis of nine distinct hsp70:lacZ fusion reporter constructs in multiple independent transgenic lines. hsp70 sequences -194 to +276 supported an average 2.7-fold increase during aging. This increase was reduced or eliminated by deletion or point mutation of the heat shock response elements, consistent with a transcriptional mechanism. Similar sequence requirements were observed for increased expression in response to catalase null mutation as a model of oxidative stress. hsp70 5'UTR sequences were required for efficient basal expression of transgenes, but were not sufficient to confer detectable upregulation during aging. Inclusion of additional hsp70 coding region sequences from +276 to + 1011 created a larger hsp70:lacZ fusion protein and had two effects: dramatic reduction of the overall expression level of the fusion protein, and an additional three to fourfold upregulation during aging. These results suggest that the coding region sequences reduce fusion protein abundance and that this negative effect decreases as a function of age. The data support a model for increased expression of hsp70 transgenes during aging involving both transcriptional and posttranscriptional components.

Aging↗