A split promoter for a eucaryotic tRNA gene

Hofstetter, H.; Kressmann, A.; Birnstiel, Max L.

doi:10.1016/0092-8674(81)90348-2

Cited by 248 publications

(163 citation statements)

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“…In the VAI RNA gene of adenovirus an internal control region, approximately 60 bp long, could be identified (3,4). In the case oftRNA genes a more precise analysis leads to the identification of two sequences of about 10 nucleotides each, located within the coding region, whose presence is essential for transcription (5,6). This overlap of coding sequences and transcriptional signals is characteristic of eukaryotic organisms: the corresponding genes (5S RNA, tRNA) in prokaryotes have a classical promoter clearly separated from the coding region and located in the 5' flanking region (7).…”

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“…We discarded the alternative of coinjecting a purified 5S RNA gene, as was done by Hofstetter et al. (5), because we found that there is a considerable degree of competition with tRNA genes (15).…”

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“…On the basis of the analysis of several mutants of a tRNATYr gene of yeast Koski et al (10) have hypothesized that the internal promoter of a tRNA gene is essentially constituted by these invariant nucleotides. It is interesting that these invariant nucleotides are particularly, although not exclusively, concentrated in the two essential regions that constitute the split promoter of a tDNAPro from Caenorhabditis elegans (6) and of a tDNAMet from Xenopus laevis (5).…”

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Relationship between the two components of the split promoter of eukaryotic tRNA genes.

Ciliberto¹,

Traboni²,

Cortese³

1982

Proc. Natl. Acad. Sci. U.S.A.

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Relationship between the two components of the split promoter of eukaryotic tRNA genes.

Ciliberto¹,

Traboni²,

Cortese³

1982

Proc. Natl. Acad. Sci. U.S.A.

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“…Region c is contained in segment C. (18) concluded that the sequence between the two essential regions, equivalent to our regions a and c, has only a "spacer" function. However, it must be stressed that because of the difference of the sequences of DNA coding for the tRNAPro and tRNAMet genes, the restriction enzyme dissection and the construction of subclones were necessarily different in the two cases.…”

Section: Bamhi (C-c-g-g-a-t-c-c-g-g) and Hindhi (C-c-a-a-g-c-t-t-g-g)mentioning

confidence: 99%

Promoter of a eukaryotic tRNAPro gene is composed of tree noncontiguous regions.

Ciliberto

Castagnoli²,

Melton³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

118

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The 71-base-pair coding sequence ofthe tRNAP1O gene from Caenorhabditis elegans contains all of the information required for transcription and processing in the injected oocytes.Several subclones of the DNA coding for the tRNAP"' were constructed, carrying deletions or insertions, or both. Their transcriptional properties lead to the hypothesis that the tRNAPW gene promoter is composed of three discontinuous regions within the coding sequence.In previous studies we have characterized steps in the synthesis of mature tRNA molecules in eukaryotic cells by injecting cloned DNAs coding for tRNA into Xenopus laevis oocytes (1-4). These studies have shown that tRNA genes are transcribed into precursor molecules, which are converted into tRNAs through a series of nucleolytic cleavages and nucleotide modifications.In order to gain further insight into the process of tRNA biosynthesis and to determine how cells regulate the expression of these genes, it will be instructive to identify the transcriptional signals present on the DNA sequence and recognized by the RNA polymerase III. The work ofBrown and his colleagues (5, 6) has revealed that the relatively small 5S RNA gene contains a 30-base-pair region within the coding sequence that is essential for initiation of transcription by RNA polymerase III. Also for tRNA genes (7-10), it appears probable that their promoters are not located in the 5' flanking region, even though this region may play a regulatory role in some cases (9, 10). It is unclear, however, whether they have an internal control region analogous to that of the 5S RNA genes. To establish this, we constructed deletion and insertion mutants, altering sequences within the coding regions of the tRNAP1O gene of Caenorhabditis elegans. This allowed the identification ofthree separated regions within the coding sequence that are important for transcription of the tRNAPr' gene. MATERIALS AND METHODSChemicals and Enzymes. Isopropyl-D-thiogalactopyranoside and 5-bromo-4-chloro-3-indolyl-D-galactoside were purchased from Sigma, and recombinant DNA linkers were purchased from Collaborative Research (Waltham, MA). T4 DNA ligase, polynucleotide kinase, and endonucleases EcoRI and Hae III were gifts of V. Pirrotta. Endonucleases HindIII and Sin I and DNA polymerase large fragment Klenow enzyme were purchased from Boehringer. Endonucleases HinfI and BamHI and nucleases S1 and BAL 31 were purchased from Bethesda Research Laboratories. Exonuclease Xma was a gift of M. Zabeau. All radioactive compounds were purchased from Amersham Buchler, Braunschweig.Bacterial Strains, Plasmids, and Cloning Vehicles. Escherichia coli K-12 (strain 71-18) was used for transformation with phage M13 (strain mp2) replicative form I and all of its recombinant derivatives (4, 11, 12). E. coli K-12 (strain K514) was used for transformation with pBR322 plasmid and its recombinantderivatives Protocolsfortransformation and preparation of double-stranded DNA were as described (1, 4).Polyacrylamide Gel Electrophoresis. DNA restriction fragment...

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“…A unique feature of the mechanism of Pol III transcription is that the signals on the DNA required for transcription initiation are located within the coding portion of the gene (2)(3)(4)(5)(6)(7)(8). This is different from the transcription mechanism of prokaryotic genes where the promoter sequence lies at the 5' side of the coding sequence and is never transcribed into RNA (9).…”

Section: Introductionmentioning

confidence: 99%