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Query: EC:2.7.7.6 (
RNA polymerase
)
34,946
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Promoters of genes for bacteriophage lambda and for Escherichia coli ribosomal RNA (rrnB), elongation factor Tu (tufB), ribosomal proteins L11 (rplK), L1 (rplA), L10 (rplJ), and L7/
L12
(rplL), and
RNA polymerase
subunits beta (rpoB) and beta' (rpoC) were studied by use of two types of filter binding assays which measured E. coli
RNA polymerase
binding and initiation of transcription on restriction fragments of lambda rifd 18 DNA. The DNA fragments selectively retained on filters were eluted, concentrated, and analyzed by gel electrophoresis. The binding characteristics of these promotor fragments were qualitatively determined by varying the
RNA polymerase
, salt, and glycerol concentrations in the polymerase binding assay with HaeIII fragments of lambda rifd 18 DNA. The approximate map locations of these small HaeIII fragments were determined by HaeIII digestion of the larger, previously mapped EcoRI, HindIII, and SmaI restriction fragments of the phage DNA. The base compositions proximal to the 5' ends of mRNA's from promoters on these DNA fragments were elucidated by the polymerase initiation assay, in which the addition of various combinations of nucleoside triphosphates to the reaction allowed
RNA polymerase
to form high-salt-resistant initiation complexes with some of the known SmaI + EcoRI, EcoRI + HindIII, or HaeIII restriction fragments of lambda rifd 18 DNA. The data obtained by this technique are consistent with the map positions and 5' mRNA base sequences of the known lambda promotors p'R, po, pR and pL. In the main focus of this work, we have determined the approximate map locations and 5' mRNA base compositions of several promoters for known E. coli genes including rrnB, tufB, rplK,A, and rplJ,L. No promoter was detected between rplL and the rpoB,C genes. Thus our data are consistent with the conclusion of Yamamoto and Nomura (1978) that the beta and beta' mRNA is probably cotranscribed from the promoter for rplJ,L. Finally, the approximate map positions and the NTP combinations which initiated transcription of several unknown lambda and E. coli in vitro promoters are reported. The methods reported should prove useful for studying the characteristics of promoters on other cloned DNA regions.
...
PMID:Escherichia coli RNA polymerase binding and initiation of transcription on fragments of lambda rifd 18 DNA containing promoters for lambda genes and for rrnB, tufB, rplC,A, rplJ,L, and rpoB,C genes. 15 6
The in vitro synthesis of elongation factor (EF)-Tu (tufB), the beta beta' subunits of
RNA polymerase
, ribosomal proteins L10 and
L12
directed by DNA from the transducing phage lambda rifd 18, EF-Tu (tufA), EF-G, and the alpha subunit of
RNA polymerase
directed by DNA from the transducing phage lambda fus3 has been investigated in a crude and a partially defined protein-synthesizing system. Proteins L10 and
L12
are synthesized in the partially defined system almost as well as in the crude system. However, the synthesis of EF-Tu, EF-G, and the alpha and beta beta' subunits of
RNA polymerase
is far less efficient in the partially defined system. An active fraction that stimulates the synthesis of these latter proteins has been obtained by fractionation of a high-speed supernatant on DEAE-cellulose. Because previous studies showed that this fraction (1 M DEAE salt eluate) contains a protein, called L factor, that stimulates beta-galactosidase synthesis in vitro, L factor was tested for activity. Although L factor stimulates the synthesis of the beta beta' subunits, it has little or no effect on the in vitro synthesis of the other products studied. In the present experiments, the ratio of
L12
/L10 and of EF-Tu (tufA)/EF-G formed is 4-6. These values are consistent with in vivo results.
...
PMID:DNA-directed in vitro synthesis of proteins involved in bacterial transcription and translation. 16 May 61
The DNA of the transducing phage lambdarifd18 contains, among others, the genes for the ribosomal proteins L11, L1, L10, and
L12
and the beta and beta' subunits of
RNA polymerase
(nucleosidetriphosphate:
RNA nucleotidyltransferase
,
EC 2.7.7.6
). In a coupled in vitro protein-synthesis system, lambdarifd18 DNA directs the synthesis of about four to five molecules of
L12
per molecule of L10. This is consistent with the finding that there are four copies of
L12
per ribosome. The ratio of
L12
/L10 was also examined from an EcoRI fragment of lambdarifd18 that contains the L10 gene and about 50% of the
L12
gene. A significantly lower ratio of truncated
L12
/L10 was observed compared to the intact phage. The binding of
RNA polymerase
to various lambdarifd18 DNA restriction fragments was used to locate possible promoter sites. These binding experiments suggest that the beta and beta' subunits of
RNA polymerase
are cotranscribed with at least ribosomal protein L12 and, also, that there may be an additional promoter site for the
L12
gene within the structural gene for L10.
...
PMID:In vitro regulation of DNA-dependent synthesis of Escherichia coli ribosomal protein L12. 28 12
The lambdarifd18 transducing phage carries genes for
RNA polymerase
(nucleosidetriphosphate:
RNA nucleotidyltransferase
;
EC 2.7.7.6
) subunits beta and beta' (rpoB,C) and genes for four ribosomal proteins (rplK for L11, rplA for LI, rplJ for L10, and rplL for L7/
L12
). DNA segments of various sizes, which cover the rifd allele of the rpoB gene, were cloned into lambda vector phages. The hybrid phages were then analyzed for their ability to express the rpoB gene and neighboring ribosomal protein genes in ultraviolet-irradiated lambda-lysogenic and nonlysogenic bacterial hosts. The results show that the rpoB gene is cotranscribed with two neighboring ribosomal protein genes and that in the rpoB,C transcription unit is: promoter, rp1J, rp1L, rpoB, and rpoC.
...
PMID:Contranscription of genes for RNA polymerase subunits beta and beta' with genes for ribosomal proteins in Escherichia coli. 35 3
The 3072-nucleotide-long sequence of a segment from the 88-min region of the Escherichia coli chromosome has been determined. The sequence covers the genes for ribosomal proteins L11 (rplK), LI (rplA), L10 (rplJ), and L7/
L12
((rplL), and the 5' end of the gene for the beta subunit of
RNA polymerase
(rpoB), along with the presumed regulatory regions for these genes. The probable locations of the promoter for the first two genes (the L11 operon) and the promoter for the latter three genes (the proximal part of the beta operon) have been identified. We have also found that the four ribosomal protein genes preferentially use codons that are recognized efficiently by the most abundant tRNA species. These and other features of the sequence results are discussed in relation to available information obtained from both in vitro and in vivo experiments on the expression of these ribosomal and RNA polymerase subunit genes.
...
PMID:Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit beta in Escherichia coli. 37 81
Fragments of lambda drifd 18 DNA with different end-points within the set of structural genes of ribosomal proteins L11 (RPLK), Li (rplA), L10 (rplJ) and
L12
(rplL) as well as the beta (rpoB) ANd beta' (rpoC) subunits of
RNA polymerase
have been cloned on plasmids. These plasmids were transformed in host cells which were mutant for each of the genes, enabling expression of both wild-type (plasmid-borne) and mutant (chromosomal) genes to be differentiated. On the basis of these results we propose the following genetic structure for the region: rplK and rplA are in one operon; rplL, rpoB and rpoC are in a second. Our data suggest the possibility that rplJ is by itself in an operon situated between the other two.
...
PMID:Expression of Escherichia coli ribosomal protein and RNA polymerase genes cloned on plasmids. 38 41
The aphid Schizaphis graminum is dependent on an association with a prokaryotic endosymbiont (Buchnera aphidicola). The nucleotide (nt) sequence of a 5040 base pair (bp) DNA fragment of B. aphidicola, homologous to the rplL-rpoB-rpoC portion of the Escherichia coli beta operon, was determined. The DNA coded for the terminal 35 amino acids of RplL (large ribosomal subunit protein L7/
L12
), the complete RpoB (beta-subunit of
RNA polymerase
), and the first 209 amino acids of RpoC (beta'-subunit of
RNA polymerase
). The deduced sequences of B. aphidicola RplL, RpoB, and RpoC were 71, 84, and 91% identical, respectively, to the homologous proteins of E. coli. The sequences of two portions of the intergenic region between rplL and rpoB were nearly identical in both B. aphidicola and E. coli. One sequence constituted an inverted repeat that could be an RNase III-messenger RNA processing site; the other sequence preceded RpoB. A compilation of the codon usage for RpoB, RpoC, and other B. aphidicola proteins indicated a major preference for A or T in the first and third positions, a result consistent with the low guanine plus cytosine (G + C) content of the DNA of this organism.
...
PMID:Sequence analysis of an aphid endosymbiont DNA fragment containing rpoB (beta-subunit of RNA polymerase) and portions of rplL and rpoC. 136 99
Using segments of the Escherichia coli rpoB and rpoC genes as heterologous probes, we have identified and cloned an 8.3 kb PstI fragment from the Staphylococcus aureus genome containing the rpoB and rpoC genes, which respectively encode the beta and beta' subunits of
DNA-directed RNA polymerase
. This region is almost certainly equivalent to the rif locus, located near to fus at interval 12/13 on the S. aureus linkage map. Limited DNA sequencing revealed the gene order rpoB-rpoC (transcribed from left to right) and identified the rplL gene, encoding ribosomal protein L7/
L12
, upstream of rpoB. This and other evidence suggests that the rpoBC genes of S. aureus form part of a large gene cluster encoding components of the transcription and translation apparatus which is well-conserved in other eubacteria.
...
PMID:Cloning and physical mapping of the Staphylococcus aureus rplL, rpoB and rpoC genes, encoding ribosomal protein L7/L12 and RNA polymerase subunits beta and beta'. 140 88
A 5789-nucleotide-long EcoRI fragment from the genome of Thermotoga maritima, identified by cross-hybridization to L11, L1, L10, and
L12
ribosomal protein gene sequences from Escherichia coli, was cloned and sequenced. The fragment encodes five tRNAs (tRNA(met1), anticodon complementary to AUG; tRNA(met2), AUG; tRNA(thr), ACA; tRNA(tyr), UAC; tRNA(trp), UGG), the transcription termination-antitermination factor nusG, the four 50 S subunit ribosomal proteins L11, L1, L10, and
L12
, and the amino-terminal portion of the
RNA polymerase
beta subunit protein. The five tRNA genes, the nusG gene, and the L11, L1, L10, and
L12
ribosomal protein genes form a complex transcription unit. Transcripts appear to be initiated from an upstream promoter, P1, located in front of the tRNA(met1) gene and from three internal promoters: P2 is located immediately in front of the tRNA(met2) gene; PL10 is near the beginning of the L1-L10 intergenic space, and PL12 is at the end of the L10 gene sequence. The tRNA sequences are excised from the leader regions of the P1- and P2-initiated transcripts. Three putative but potentially important regulatory sequences were identified within this operon: an L1 translational control site, a transcription attenuator, and a strong rho-independent terminator. The strong terminator located distal to the
L12
gene overlaps a fifth promoter, P beta, which is used to initiate transcripts of the downstream
RNA polymerase
beta subunit gene. The T. maritima NusG protein exhibits 43% amino acid sequence identity when aligned to the E. coli protein; the alignment is interrupted by a large 171-amino acid-long insertion into the T. maritima protein after codon 45.
...
PMID:The organization and expression of essential transcription translation component genes in the extremely thermophilic eubacterium Thermotoga maritima. 142 27
The relationship between global RNA transcription capacity and transcript initiation, attenuation, and stability in the rplKAJLrpoBC operon of Escherichia coli has been examined. The rplKAJLrpoBC operon encodes in order the four large ribosome subunit proteins, L11, L1, L10, and
L12
, and the two large beta and beta' subunits of
RNA polymerase
. Operon transcripts are initiated at two promoters, PL11 and PL10. The
L12
-beta intergenic space contains a transcription attenuator which, during balanced growth, terminates about 80% of the transcripts exiting the
L12
gene; the remaining transcripts read through into the beta and beta' encoding genes. The capacity for global transcription initiation was modulated using a strain carrying a temperature-sensitive, initiation-defective mutation in rpoC. Following a shift to 39 degrees C, the global transcription initiation capacity was reduced to about one-half the level at 30 degrees C. This partial restriction resulted in a decrease in the stability of distal beta mRNA, whereas the stability of proximal L11-L1 and L10-
L12
mRNA was not changed. Measurements of the synthesis rates of L11-L1, L10-
L12
, and beta mRNAs relative to total RNA synthesis indicated that this operon was selectively transcribed when the initiation capacity of
RNA polymerase
was limited. The synthesis rates of L11-L1 and L10-
L12
mRNA increased about 2-fold, whereas the synthesis rate of beta mRNA increased nearly 5-fold. The relative transcription of other ribosome component genes and the alpha subunit gene exhibited only a modest increase during the partial restriction. Protection from S1 nuclease was used to demonstrate that the preferential transcription within the operon of beta mRNA was the consequence of active regulation of termination-antitermination at the attenuator structure in the
L12
-beta intergenic space. These results demonstrate that global transcription capacity may be an important parameter in determining both initiation and attenuation of transcription of the rplKAJLrpoBC ribosomal protein-
RNA polymerase
operon.
...
PMID:RNA polymerase activity may regulate transcription initiation and attenuation in the rplKAJLrpoBC operon in Escherichia coli. 198 49
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