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W L Gerlach

Publications and source records attributed to W L Gerlach.

At least 37 records · Page 2Linked to original sources
PubMed1989

Sequences required for self-catalysed cleavage of the satellite RNA of tobacco ringspot virus.

The satellite RNA of tobacco ringspot virus (sTobRV) undergoes self-catalysed cleavage during replication. A plasmid for in vitro expression of sTobRV has been constructed and used to obtain a library of mutagenized sTobRV sequences. Screening of these mutants has allowed precise definition of the sequences required for (+) and (-) strand cleavage. The sequences and RNA structures associated with cleavage of each strand differ markedly. Cleavage of the (+) strand requires those sequences flanking the site for cleavage to form a 'hammerhead' domain, similar to those found in other satellite and viroid RNA. In contrast, cleavage of the (-) strand requires only a small region of 12 nucleotides (nt) at the site of cleavage, and a sequence of 55 nt positioned elsewhere in the molecule. Comparison with a closely related satellite suggests that a novel RNA structure may be involved in (-) strand cleavage.

Base Sequence↗
PubMed1988

Anaerobically regulated aldolase gene of maize. A chimaeric origin?

The sequence of the anaerobically induced fructose 1,6-bisphosphate aldolase gene of maize is presented. Analysis of the upstream sequences of the aldolase gene reveals a six base-pair sequence (TGGTTT) with perfect homology to one of the sub-regions of the anaerobic regulatory element (ARE) which is responsible for the anaerobic induction of the maize alcohol dehydrogenase 1 gene (Adh1). In the aldolase gene this sequence is located at position -70 relative to the start of transcription, in a small segment proven by functional analysis to be important for expression of the aldolase gene. Since this six base-pair sequence has been shown to be critical for anaerobic induction of the Adh1 mRNA, is in the functional promoter region of aldolase and is also present in a homologous position in Adh2 (another anaerobically-induced gene), we suggest this hexanucleotide is essential for anaerobic regulation of each of these genes. The maize aldolase gene is about 50% homologous at the amino acid level to the animal aldolase gene but has a completely different intron/exon structure. While the rat aldolase gene has nine introns the maize gene has a single large intron near the N terminus of the coding region. Because there is 55% homology downstream from the intron and very little homology upstream, we suggest that the maize gene has acquired a 5' region containing signals for anaerobic regulation and fortuitously adding a new N-terminal region to the protein. We must suppose that the plant gene has lost the remaining introns.

Anaerobiosis↗
PubMed1988

Simple RNA enzymes with new and highly specific endoribonuclease activities.

In vitro mutagenesis of sequences required for the self-catalysed cleavage of a plant virus satellite RNA has allowed definition of an RNA segment with endoribonuclease activity. General rules have been deduced for the design of new RNA enzymes capable of highly specific RNA cleavage, and have been successfully tested against a new target sequence.

Arginine↗
PubMed1988

Sequence and organization of barley yellow dwarf virus genomic RNA.

The nucleotide sequence of the genomic RNA of barley yellow dwarf virus, PAV serotype was determined, except for the 5'-terminal base, and its genome organization deduced. The 5,677 nucleotide genome contains five large open reading frames (ORFs). The genes for the coat protein (1) and the putative viral RNA-dependent RNA polymerase were identified. The latter shows a striking degree of similarity to that of carnation mottle virus (CarMV). By comparison with corona- and retrovirus RNAs, it is proposed that a translational frameshift is involved in expression of the polymerase. An ORF encoding an Mr 49,797 protein (50K ORF) may be translated by in-frame readthrough of the coat protein stop codon. The coat protein, an overlapping 17K ORF, and a 3'6.7K ORF are likely to be expressed via subgenomic mRNAs.

Amino Acid Sequence↗
PubMed1988

The Ds1 transposable element acts as an intron in the mutant allele Adh1-Fm335 and is spliced from the message.

The Ds-induced maize Adh1 allele Adh1-Fm335 retains its anaerobic regulation and normal transcription start site despite the presence of the 405 bp Ds element in the 5' untranslated leader region of the gene. The steady state level of Adh1-specific transcript is reduced to about 1% that of the progenitor or revertant alleles. Run-on transcription studies show that the reduced level of Adh1 specific mRNA is not attributable to a decreased transcription rate. S1 mapping indicates that the Ds element is spliced from the Adh1-Fm335 transcript using a donor site 14 bp into the Ds element and an acceptor site at the 3' junction of the Ds element with the flanking genome DNA.

Alcohol Dehydrogenase↗
PubMed1988

Sequence and identification of the barley yellow dwarf virus coat protein gene.

The nucleotide sequence of the coat protein gene of barley yellow dwarf virus (BYDV, PAV serotype) was determined, and the amino acid sequence was deduced. The open reading frame, encoding a protein of relative molecular mass (Mr) 22,047, was confirmed as the coat protein gene by comparison with amino acid sequences of tryptic peptides derived from dissociated virions. In addition, a fragment of this gene expressed in Escherichia coli produced a product which was recognized by antibodies prepared against purified BYDV virions. An overlapping reading frame encoding an Mr 17,147 protein is contained completely within the coat protein gene.

Amino Acid Sequence↗
PubMed1987

The Ds1 controlling element family in maize and Tripsacum.

Hybridization experiments indicated that the maize genome contains a family of sequences closely related to the Ds1 element originally characterized from the Adh1-Fm335 allele of maize. Examples of these Ds1-related segments were cloned and sequenced. They also had the structural properties of mobile genetic elements, i.e., similar length and internal sequence homology with Ds1, 10- or 11-bp terminal inverted repeats, and characteristic duplications of flanking genomic DNA. All sequences with 11-bp terminal inverted repeats were flanked by 8-bp duplications, but the duplication flanking one sequence with 10-bp inverted repeats was only 6 bp. Similar Ds1-related sequences were cloned from Tripsacum dactyloides. They showed no more divergence from the maize sequences than the individual maize sequences showed when compared with each other. No consensus sequence was evident for the sites at which these sequences had inserted in genomic DNA.

Base Sequence↗
PubMed1986

Satellite tobacco ringspot virus RNA: A subset of the RNA sequence is sufficient for autolytic processing.

The satellite RNA of tobacco ringspot virus depends upon tobacco ringspot virus for its replication and source of coat protein. The satellite RNA reduces virus accumulation and the severity of virus-induced symptoms. Repetitive sequence, dimeric, and higher forms of the satellite RNA are known to autolytically process to form biologically active monomeric RNA of 359 nucleotide residues [Prody, G. A., Bakos, J. T., Buzayan, J. M., Schneider, I. R. & Bruening, G. (1986) Science 231, 1577-1580], with a 5'-hydroxyl and a 2',3'-cyclic phosphodiester as the new terminal groups. We show here that transcripts of full-length and truncated DNA clones of the satellite RNA sequence also process in a nonenzymic reaction. One such transcript was an RNA that has about one-fourth of the satellite RNA sequence, representing the 3'-terminal and 5'-terminal portions of monomeric RNA joined in the junction that is cleaved in dimeric RNA. This RNA autolytically processed more efficiently than molecules with a larger proportion of the satellite RNA nucleotide sequence.

Journal Article↗
PubMed1985

Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize.

The Adh2 gene of maize has a nucleotide sequence closely related to that of the maize Adh1 gene indicating that the two genes arose from a progenitor gene by a duplication event. The coding regions are 82% conserved at the nucleotide level and 87% conserved at the amino acid level. Each gene has nine introns in identical positions but their nucleotide sequences and lengths differ. Adh2 encodes a single mRNA and has a possible polyadenylation signal, AATAAT, fifteen bases upstream from the polyadenylation site. The Adh2 gene together with the Adh1 gene is induced by anaerobic conditions and under these conditions produces an increased level of mRNA. The 5' untranslated regions of the transcripts of Adh1 (100 bp) and Adh2 (126 bp) have diverged in sequence except for a conserved region which may be important for their anaerobic-specific translation. The 3' and 5' flanking sequences of the two genes are also divergent except for 11 bp of precise homology around the TATA boxes and three other 8 bp sequences further upstream. One of the common 8 bp sequences (CACCTCCC) is in a similar position (-150 bp to -200 bp) in the two genes and may have a regulatory function in gene expression.

Alcohol Dehydrogenase↗
PubMed1984

Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize.

A cDNA clone of maize Adh1 which contains the entire protein coding region of the gene has been constructed. The protein sequence predicted from the nucleotide sequence is in agreement with limited protein sequencing data for the ADH1 enzyme. An 11.5 kb genomic fragment containing the Adh1 gene has been isolated using the cDNA clone as a probe, and the gene region fully sequenced. The gene is interrupted by 9 introns, their junction sequences fitting the animal gene consensus sequence. Within the gene there is a triplication of a segment (104 bp) spanning an intron-exon junction. Presumptive promoter elements have been identified and are similar in nucleotide sequence and location, relative to the start of transcription, to those of other plant and animal genes. No recognizable poly(A+) addition signal is evident. Comparison of the nucleotide sequences of the cDNA (derived from an Adh1 -F allele) and genomic (derived from an Adh1 -S allele) clones has identified an amino acid difference consistent with the observed difference in electrophoretic mobility of the two enzymes. The maize ADH1 amino acid sequence is 50% homologous to that of horse liver ADH but is only 20% homologous to yeast ADH.

Alcohol Dehydrogenase↗
PubMed1982

cDNA cloning and induction of the alcohol dehydrogenase gene (Adh1) of maize.

cDNA clones of Adh1, one of two genes encoding alcohol dehydrogenase (ADH; alcohol:NAD(+) oxidoreductase, EC 1.1.1.1) in the maize genome, have been isolated. They were derived from mRNA extracted from anaerobically treated roots of maize seedlings. Identification was initially made on the basis of molecular weight and electrophoretic properties of the in vitro polypeptide obtained in hybridization-release-translation experiments. The identification was confirmed by antibody precipitation and by the use of maize stocks having different genetic constitutions at the Adh1 locus. The sequence of the longest cDNA segment, approximately 900 base pairs, was determined and appears to code for 168 COOH-terminal amino acids and to have a 3' nontranslated region of 364 base pairs. Reverse Southern hybridizations established that two different Adh1-S stocks produce a mRNA of 1,650 nucleotides, whereas an additional mRNA of 1,750 nucleotides is produced in three Adh1-F stocks. A 50-fold increase in Adh1 mRNA level occurs during anaerobiosis, reaching a maximum at 5 hr. Return to aerobic conditions indicates a half-life of more than 18 hr for the anaerobically induced Adh1 mRNA.

Journal Article↗
PubMed1980

Sequence organization of the repeating units in the nucleus of wheat which contain 5S rRNA genes.

Examples of both the 410 and 500 bp size classes of repeating units containing wheat 5S rRNA genes have been cloned in plasmid pBR322 and sequenced. The structural genes showed sequence microheterogeneity. Also the gene in the 500 bp repeat which was sequenced had a 15 bp tandem duplication within it and appears to be representative of non-transcribed subfamily of repeating units. The transcription terminators comprise 14-17 A.T bp immediately preceded by a region of weak dyad symmetry. The spacer regions adjacent to the transcription terminators in the two different size repeat units have interspersed oligonucleotides of high and low homology. The central spacer regions of the two size classes have very different sequences. The only repeated sequence in the spacers has undergone extensive divergence. In contrast to most of the spacer, the 70 bp region preceding the genes of each type of repeat show high homology, suggesting that it has functional importance. The transcription start point obeys the pyrimidine-1 purine+1 rule.

Base Sequence↗
PubMed1979

Cloning and characterization of ribosomal RNA genes from wheat and barley.

Wheat and barley DNA enriched for ribosomal RNA genes was isolated from actinomycin D-CsCl gradients and used to clone the ribosomal repeating units in the plasmid pAC184. All five chimeric plasmids isolated which contained wheat rDNA and eleven of the thirteen which had barley rDNA were stable and included full length ribosomal repeating units. Physical maps of all length variants cloned have been constructed using the restriction endonucleases Eco Rl, Bam Hl, Bgl II, Hind III and Sal I. Length variation in the repeat units was attributed to differences in the spacer regions. Comparison of Hae III and Hpa II digestion of cereal rDNAs and the cloned repeats suggests that most methylated cytosines in natural rDNA are in -CpG-. Incomplete methylation occurs at specific Bam Hl sites in barley DNA. Detectable quantities of ribosomal spacer sequences are not present at any genomic locations other than those of the ribosomal RNA gene repeats.

Base Sequence↗
PubMed1975

Genetic properties of some amber-ochre supersuppressors in Saccharomyces cerevisiae.

21 amber-ochre supersuppressor alleles have been isolated in a strain of Saccharomyces cerevisiae. Their dominance-recessiveness, temperature sensitivity, allelism, intergenic and intragenic complementation properties and their effect on cell viability and colony growth rate have been characterized. They are compared with the yeast amber-ochre supersuppressors described by Inge-Vechtomov and Andrianova (1970a, b) and Hawthorne and Leupold (1974). The possible molecular basis of their suppressor activity is discussed in relation to their genetic properties.

Alleles↗