Biomaterial Database

Three regions of a yeast tRNALeu3 gene promote RNA polymerase III transcription. 1984

J D Johnson, and G J Raymond

A Saccharomyces cerevisiae tRNALeu3 gene has been dissected to identify sequences essential for recognition by the yeast RNA polymerase III transcription apparatus. Three putative promoter regions have been identified, one each in the 3'- and 5'-halves of the tRNA coding sequence, the A- and B-blocks, respectively, and one in the 5'-flanking region. DNA fragments derived from the intact gene and bearing the 5'-flanking region, the 5'-flanking region plus the A-block, the A- and B-blocks without the 5'-flanking region, the 5'-flanking region with the B-block, and the B-block only have been subcloned. Plasmids carrying these fragments were used as templates in a homologous in vitro transcription assay to determine the contributions of the various sequences to the template activity of the gene. No template-dependent transcription was seen when fragments with only the 5'-flanking region or B-block were tested. A very weak template-dependent transcript was observed from clones bearing the A- and B-block regions. Transcription of the fragment bearing the 5'-flanking sequence and A-block was considerably more efficient but reduced relative to the intact gene. The clone including the 5'-flanking region and 3'-half of the gene is transcribed by the yeast extract with an efficiency approaching that of the intact gene. Partial deletions were constructed in which the highly conserved 5'-flanking pentadecanucleotide sequence was replaced by vector DNA. Replacement of the sequence between positions -12 and -2 (relative to the tRNA coding sequence) decreased transcription efficiency 10-fold even though the A- and B-blocks were left intact. We conclude that this 5'-flanking region, in conjunction with either the A- or B-block sequence is sufficient to constitute a promoter for the yeast RNA polymerase III transcription apparatus.

UI	MeSH Term	Description	Entries
D009154	Mutation	Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.	Mutations
D010957	Plasmids	Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.	Episomes,Episome,Plasmid
D002872	Chromosome Deletion	Actual loss of portion of a chromosome.	Monosomy, Partial,Partial Monosomy,Deletion, Chromosome,Deletions, Chromosome,Monosomies, Partial,Partial Monosomies
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D004271	DNA, Fungal	Deoxyribonucleic acid that makes up the genetic material of fungi.	Fungal DNA
D005800	Genes, Fungal	The functional hereditary units of FUNGI.	Fungal Genes,Fungal Gene,Gene, Fungal
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D012320	RNA Polymerase III	A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure where it transcribes DNA into RNA. It has specific requirements for cations and salt and has shown an intermediate sensitivity to alpha-amanitin in comparison to RNA polymerase I and II.	DNA-Dependent RNA Polymerase III,RNA Polymerase C,DNA Dependent RNA Polymerase III,Polymerase C, RNA,Polymerase III, RNA
D012321	DNA-Directed RNA Polymerases	Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).	DNA-Dependent RNA Polymerases,RNA Polymerases,Transcriptases,DNA-Directed RNA Polymerase,RNA Polymerase,Transcriptase,DNA Dependent RNA Polymerases,DNA Directed RNA Polymerase,DNA Directed RNA Polymerases,Polymerase, DNA-Directed RNA,Polymerase, RNA,Polymerases, DNA-Dependent RNA,Polymerases, DNA-Directed RNA,Polymerases, RNA,RNA Polymerase, DNA-Directed,RNA Polymerases, DNA-Dependent,RNA Polymerases, DNA-Directed
D012346	RNA, Transfer, Amino Acyl	Intermediates in protein biosynthesis. The compounds are formed from amino acids, ATP and transfer RNA, a reaction catalyzed by aminoacyl tRNA synthetase. They are key compounds in the genetic translation process.	Amino Acyl tRNA,Transfer RNA, Amino Acyl,tRNA-Amino Acyl,Amino Acyl T RNA,Acyl tRNA, Amino,Acyl, tRNA-Amino,tRNA Amino Acyl,tRNA, Amino Acyl