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Query: UNIPROT:P06889 (
Mol
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630,302
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Inverted sequences of the chloramphenicol acetyltransferase (CAT) reporter gene were fused to a soybean tRNA(met(i)) gene lacking a terminator such that the tRNA(met(i)) sequences caused the co-transcription of CAT antisense sequences by
RNA polymerase III
. When electroporated into carrot protoplasts, these antisense DNA constructs suppressed CAT enzyme activity expressed from co-electroporated DNAs containing the CAT gene downstream of the cauliflower mosaic virus (CaMV) 35S RNA promoter. Our most effective construct, an antisense sequence complementary to the 3' portion of the CAT gene, inhibited CAT activity five-fold greater than an antisense construct expressed by RNA polymerase II from the cauliflower mosaic virus 35S RNA promoter. These results indicate that antisense sequences transcribed by
RNA polymerase III
should efficiently suppress gene expression in plants.
Plant
Mol
Biol 1992 Jul
PMID:Suppression of gene expression in plant cells utilizing antisense sequences transcribed by RNA polymerase III. 162 77
We report here the isolation, sequence analysis, structure, and expression of the gene encoding the largest subunit of
RNA polymerase III
(RPIII) from Plasmodium falciparum. The P. falciparum RPIII gene consists of 5 exons and 4 introns, is expressed in all of the asexual erythrocytic stages of the parasite as a 8.5-kb mRNA, and is present in a single copy on chromosome 13. The predicted 2339 amino acid residue RPIII subunit contained 5 regions that were conserved between different eukaryotic RPIII subunits, and 4 variable regions that separated the conserved regions. Three of the variable regions were greatly enlarged in comparison to the corresponding variable regions in other RPIII subunits. Variable region C' represented nearly one-third of the P. falciparum RPIII subunit (750 amino acid residues), included a unique repeated decapeptide sequence, and had some homology with yeast DNA topoisomerase II. Noteworthy amino acid sequences and structures were identified in both the conserved regions and in the enlarged variable regions, and their possible role(s) as domains that regulate RPIII enzyme activity is discussed.
Mol
Biochem Parasitol 1991 Jun
PMID:Characterization of the gene encoding the largest subunit of Plasmodium falciparum RNA polymerase III. 165 54
The ribosomal gene intergenic region from Arabidopsis thaliana contains four clusters of mutually unrelated repeated sequences. By comparison with the respective regions in two other Brassicaceae, Raphanus and Sinapis, the putative promoter sequence for
RNA polymerase I
was located. The homologies suggest that the
RNA polymerase I
promoter in Brassicaceae ranges further upstream than in animals. Upstream duplications of at least a part of the promoter region were found to be located between individual blocks of the largest internal repeat family ("A" repeats), which is made up of multiple repeats of two closely related sequences 21 or 20 bp in length. Overall structural similarities of the A. thaliana rDNA intergenic region with those from wheat and from Xenopus laevis are discussed. We also present data on the range of intraspecific length heterogeneities found in the central EcoRI fragment of the intergenic region and on the frequencies with which specific length variants occur in the genome. To determine the nature of the length heterogeneities, we sequenced the central EcoRI fragments from four independently isolated genomic clones. Three levels of rearrangements were detected. Length variation can be caused by duplication of a whole A repeat block, or, most frequently, by insertion and/or deletion of one or a few A repeat units. Surprisingly, single base mutations are extremely rare, which hints at some mechanism of homogenization which might be acting on the intergenic region. A possible function of the described sequences in transcriptional regulation is discussed and will be the aim of further investigations.
J
Mol
Biol 1991 Oct 20
PMID:rDNA intergenic region from Arabidopsis thaliana. Structural analysis, intraspecific variation and functional implications. 168 99
Choriogenic follicular cells of the silkmoth Bombyx mori contain significant quantities of antisense RNA transcribed from chorion genes. Antisense RNA derived from a chorion gene with a high content of cysteine, HcB.12, was characterized in detail. The antisense transcripts are initiated downstream from the 3' end of HcB.12 mRNA and extend over 75% of the length of the gene, comprising its entire second exon and part of its intervening sequence. The antisense RNA is devoid of any significant open reading frames and is not polyadenylated. These features, combined with the presence of specific sequence motifs within its transcribed and upstream region, suggest that antisense RNA may be transcribed by
RNA polymerase III
. Chorion antisense RNA is detectable only in choriogenic follicular cells and appears to be co-ordinately regulated with chorion mRNA. Its cytoplasmic accumulation during choriogenesis parallels that of the corresponding mRNA. Although chorion mRNA is at least five times more abundant than antisense RNA, the latter is present as a single-stranded entity in follicular cytoplasm but can form perfect duplexes with its mRNA complement upon annealing in vitro. The possible involvement of antisense RNA transcription in the pathway that controls the programmed expression of chorion genes at the level of transcription initiation or post-transcriptional processing is discussed.
J
Mol
Biol 1990 May 05
PMID:Silkmoth chorion antisense RNA. Structural characterization, developmental regulation and evolutionary conservation. 169 92
The murine gene pro1 has been cloned from JB6 epidermal cell lines that are sensitive to neoplastic transformation by tumor promoters. Insensitive JB6 variants acquire susceptibility to neoplastic transformation by tumor promoters when transfected with pro1. The repetitive nature of pro1 was indicated by sequence and Southern analysis. In contrast, northern analysis of RNA from promotion-sensitive cells revealed the presence of a small pro1-hybridizing transcript. Strand-specific RNA probes implicated an
RNA polymerase III
(RNAPIII) coding domain in pro1 as the source of this hybridization signal. Ribonuclease protection of gel-purified pro1 RNA from JB6 variant cell lines identified a 130-nucleotide transcript. The size of this transcript is compatible with in vitro RNAPIII transcription of pro1. Deletion mapping of pro1 by exonuclease III demonstrated that the biologically active domain included the RNAPIII transcription unit. RNA probes map pro1 RNA within the activity domain. These results delineate an activity domain of 597 nucleotides and suggest that a small RNA is the product of promotion-sensitivity gene pro1.
Mol
Carcinog 1990
PMID:Deletion mapping of tumor promotion-susceptibility gene pro1 implicates an RNA polymerase III transcription unit. 169 83
Structural resemblance of the human Alu family with a subset of vertebrate tRNAs was detected. Of four tRNAs, tRNA(Lys), tRNA(Ile), tRNA(Thr), and tRNA(Tyr), which comprise a structurally related family, tRNA(Lys) is the most similar to the human Alu family. Of the 76 nucleotides in lysine tRNA (including the CCA tail), 47 are similar to the human Alu family (60% identity). The secondary structure of the human Alu family corresponding to the D-stem and anticodon stem regions of the tRNA appears to be very stable. The 7SL RNA, which is a progenitor of the human Alu family, is less similar to lysine tRNA (55% identity), and the secondary structure of the 7SL RNA folded like a tRNA is less stable than that of the human Alu family folded likewise. Insertion of the tetranucleotide GAGA, which is an important region of the second promoter for
RNA polymerase III
in the Alu sequence, occurred during the deletion and ligation process to generate the Alu sequence from the parental 7SL RNA. These results suggest that the human Alu family was generated from the 7SL RNA by deletion, insertion, and mutations, which thus modified the ancestral 7SL sequence so that it could form a structure more closely resembling lysine tRNA. The similarities of several short interspersed sequences to the lysine tRNA were also examined. The Galago type 2 family, which was reported to be derived from a methionine initiator tRNA, was also found to be similar to the lysine tRNA. Thus lysine tRNA-like structures are widespread in genomes in the animal kingdom. The implications of these findings in relation to the mechanism of generation of the human Alu family and its possible functions are discussed.
J
Mol
Evol 1990 Dec
PMID:Transfer RNA-like structure of the human Alu family: implications of its generation mechanism and possible functions. 170 38
We found that hydrolysates of poly(A)RNA from Ehrlich ascites carcinoma cells which were transcribed by
RNA polymerase III
contained an unusual component designated as X. It was part of B2 RNA representing a transcript of B2 retroposon, typical of rodents. The component X possesses a cap-like structure, xPPP5'G, where x has al non-nucleotide structure. About half of all B2 RNAs contained this group at the 5'-end. Previously, Epstein et al. (Epstein P., Reddy R., Henning D., Busch H. parallel J. Biol. Chem. 1980. V. 255. P. 8901-8906) detected a similar structure at the 5'-end of small nuclear U6RNA. Later, Singh and Reddy (Singh R., Reddy R. parallel Proc. Natl. Acad. Sci. USA. 1989. V. 86. P. 8280-8283) showed methyl to be the blocking group in the component X of U6RNA. Besides B2 RNA, we found 5'-ends containing methyl groups in 7S K-RNA.
Mol
Biol (Mosk)
PMID:[B2 RNA and 7SK RNA, transcripts of RNA-polymerase III, have a cap-like structure at the 5'-end]. 171 20
Starting with two temperature-sensitive mutants (rpa190-1 and rpa190-5) of Saccharomyces cerevisiae, both of which are amino acid substitutions in the putative zinc-binding domain of the largest subunit (A190) of
RNA polymerase I
, we have isolated many independent pseudorevertants carrying extragenic suppressors (SRP) of rpa190 mutations. All the SRP mutations were dominant over the corresponding wild-type genes. They were classified into at least seven different loci by crossing each suppressed mutant with all of the other suppressed mutants and analyzing segregants. SRP mutations representing each of the seven loci were studied for their effects on other known rpa190 mutations. All of the SRP mutations were able to suppress both rpa190-1 and rpa190-5. In addition, one particular suppressor, SRP5, was found to suppress two other rpa190 mutations as well as an rpa190 deletion. Southern blot analysis combined with genetic crosses demonstrated that SRP5 maps to a region on chromosome XV loosely linked to rpa190 and represents a transposed mutant gene in two copies. Analysis of the A190 subunit by using anti-A190 antiserum indicated that the cellular concentration of A190 and hence of
RNA polymerase I
decreases in rpa190-1 mutants after a shift to 37 degrees C and that in the mutant strain carrying SRP5 this decrease is partially alleviated, presumably because of increased synthesis caused by increased gene dosage. These results suggest that the zinc-binding domain plays an important role in protein-protein interaction essential for the assembly and/or stability of the enzyme, regardless of whether it also participates directly in the interaction of the assembled enzyme with DNA.
Mol
Cell Biol 1991 Feb
PMID:Suppressor analysis of temperature-sensitive RNA polymerase I mutations in Saccharomyces cerevisiae: suppression of mutations in a zinc-binding motif by transposed mutant genes. 184 71
Cycloheximide (Cyh), administered at a dose of 5 mg/kg body wt blocks protein synthesis in normal rat liver (NRL) and regenerating rat liver (RRL). The rate of synthesis of 45S pre-rRNA in RRL, studied after RNA labelling in vivo is activated 2.8 times. Pre-r RNA synthesis in RRL is more sensitive to the stopped translation, but never falls down to the level in NRL. The major contribution to the rDNA transcription activation in RRL comes from the 20-fold increase in the number of pol I molecules engaged in the transcription, the elongation rate being 1.4-fold accelerated. Cyh quenches partially the enhanced rDNA transcription in RRL: the number of pol I molecules and their elongation rate are about 1.7-fold and 1.5-fold higher, respectively, than the corresponding values in NRL after Cyh treatment. The results show that two different mechanisms control the number and the rate of initiation and elongation of
RNA polymerase I
in rat liver; one of them depends on continuous protein synthesis and can be inactivated by Cyh, the other is Cyh resistant.
Mol
Biol Rep 1991 Feb
PMID:Activated ribosomal RNA synthesis in regenerated rat liver upon inhibition of protein synthesis. 187 19
The amino acid sequences of the largest subunits of the RNA polymerases I, II, and III from eukaryotes were compared with those of archaebacterial and eubacterial homologs, and their evolutionary relationships were analyzed in detail by a recently developed tree-making method, the likelihood method of protein phylogeny, as well as by the neighbor-joining method and the parsimony method, together with bootstrap analyses. It was shown that the best tree topologies predicted by the first two methods are identical, whereas the last one predicts a distinct tree. The maximum likelihood tree revealed that, after the separation from archaebacteria, the three eukaryotic RNA polymerases diverged from an ancestral precursor in the eukaryotic lineage. This result is contrasted with the published result showing multiple origins for the three eukaryotic polymerases. It was shown that eukaryotic
RNA polymerase I
evolved much more rapidly than RNA polymerases II and III: The N-terminal half of
RNA polymerase I
shows an extraordinarily high evolutionary rate, possibly due to relaxed functional constraints. In contrast the evolutionary rate of archaebacterial RNA polymerase is remarkably limited. In addition, including the second largest subunit of the RNA polymerase, a detailed analysis for the branching pattern of the three major groups of archaebacteria was carried out by the maximum likelihood method. It was shown that the three major groups of archaebacteria are likely to form a single cluster; that is, archaebacteria are likely to be monophyletic as originally proposed by Woese and his colleagues.
J
Mol
Evol 1991 Jan
PMID:Evolution of RNA polymerases and branching patterns of the three major groups of Archaebacteria. 190 70
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