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Biomedical subjects

A M Metz

Publications and source records attributed to A M Metz.

At least 19 recordsLinked to original sources

Two telomerase reverse transcriptases (TERTs) expressed in Candida albicans.

The human pathogenic yeast Candida albicans contains two telomerase reverse transcriptase (TERT) genes. CaTERT1 and CaTERT2 appear either to be two alleles of the same gene or two entirely different genes that encode 867-residue proteins that differ by five amino acids. Both TERTs have a calculated pI of 9.5 and a M(r) of 100.9 kDa and are the smallest TERTs identified to date. Both genes appear to be expressed. Based on sequence similarity between CaTERT1 and the Saccharomyces cerevisiae orthologue Est2p, we suggest a revised alignment for motif E of Est2p. The identification of these TERT genes provides the first opportunity to study telomerase in an important human pathogen.

Alleles↗

Updating of working memory in a running memory task: an event-related potential study.

The aim of the study was to identify central executive activity in the event-related potential (ERP) in the time and space domain. Lists of three to eight consonants were presented sequentially. After each list the ordered recall of the three most recent items was required. In this running memory task the updating of working memory contents from the fourth letter on may be understood as a control process. ERPs elicited from each consonant presented in the lists were subtracted from those of a control condition that was also applied to the participants. The difference waveforms showed fronto-central distributed positivities, probably indicating the activity of a postulated central executive. This finding confirms those of neuroimaging studies that localize executive activity to prefrontal brain areas.

Adult↗

Eukaryotic initiation factor 4B from wheat and Arabidopsis thaliana is a member of a multigene family.

Clones of eukaryotic initiation factor (eIF) 4B from wheat and Arabidopsis thaliana were obtained from cDNA and genomic libraries. The exon/intron organization of the genes from wheat and A. thaliana is very similar. The deduced amino acid sequences for the wheat and Arabidopsis eIF4B proteins showed overall similarity to each other, but very little similarity to eIF4B from other eukaryotes. The recombinant form of eIF4B supports polypeptide synthesis in an in vitro translation system and reacts with antibodies to native wheat eIF4B. In contrast to mammalian eIF4B and eIF4A, the combination of wheat eIF4B and eIF4A does not stimulate RNA-dependent ATP hydrolysis activity; however, wheat eIF4B does stimulate eIF4F and eIF4A RNA-dependent ATP hydrolysis activity. Interestingly, eIF4B does not stimulate eIF(iso)4F and eIF4A hydrolysis activity. Gel filtration experiments indicate wheat eIF4B, like its mammalian counterpart, self-associates to form a homodimer.

Amino Acid Sequence↗

Translation initiation factors eIF-iso4G and eIF-4B interact with the poly(A)-binding protein and increase its RNA binding activity.

The 5'-cap and the poly(A) tail act synergistically to increase the translational efficiency of eukaryotic mRNAs, which suggests that these two mRNA elements communicate during translation. We report here that the cap-associated eukaryotic initiation factors (eIFs), i. e. the two isoforms of the cap-binding complex (eIF-4F and eIF-iso4F) and eIF-4B, bind to the poly(A)-binding protein (PABP) both in the presence and absence of poly(A) RNA. The interactions between PABP and eIF-4F, eIF-iso4F, and eIF-4B were measured in the absence of poly(A) RNA using far Western analysis and confirmed by direct fluorescence titration studies. The functional consequence of the interaction between these initiation factors and PABP was examined using RNA binding assays and RNA mobility shift analysis. eIF-4F, eIF-iso4F, and eIF-4B promoted PABP activity through a shift in its equilibrium affinity for poly(A). eIF-iso4G, the large subunit of eIF-iso4F, was the subunit responsible for the interaction between eIF-iso4F and PABP and was the subunit that promoted PABP RNA binding activity. Truncation analysis of eIF-iso4G indicated that a domain close to its N-terminal end appeared to be involved in binding PABP. These results suggest that the interaction between PABP and eIF-4B and eIF-iso4G may be involved in mediating the functional co-dependence observed between the cap and the poly(A) tail during translation.

Binding Sites↗

Assignment of the beta-subunit of wheat eIF2 by protein and DNA sequence analysis and immunoanalysis.

Wheat germ initiation factor 2 (eIF2), like mammalian and yeast eIF2, contains three nonidentical subunits. The estimated molecular weights for the wheat subunits are 38,000 (p38), 42,000 (p42), and 50,000 (p50). Peptide sequence was obtained for the p38 subunit of wheat eIF2 and the resulting amino acid sequence suggested that it was actually the equivalent of the mammalian beta-subunit. A wheat sprout cDNA expression library was screened with antibody affinity purified to the p38 subunit. The DNA sequence of the clones obtained also indicated that the p38 subunit was the equivalent to the mammalian beta-subunit. The wheat p38 subunit was then expressed in Escherichia coli and antibodies raised to the purified recombinant protein. Only the p38 subunit of purified wheat germ eIF2 reacted with the antisera. The p38 subunit of wheat eIF2 is therefore the equivalent of mammalian eIF2beta.

Amino Acid Sequence↗

[Effect of preload of working memory on components of evoked potentials during encoding of sequentially presented letter sequences].

In recent experiments with a pseudo-random sequence of 7 consonants to recall after trial (stimulus onset asynchrony 1000 ms) a reverse relationship between P300 amplitude of the event-related potential (ERP) and letter presentation position was shown (Grune et al., 1996). It was assumed that this relationship reflects competition between encoding and retention processes in this time regime. In order to find evidence for the resource competition hypothesis in working memory a 4 x 4 grid containing 4 digits preceded a sequence of 6 consonants. In the first block subjects were instructed to ignore the grid, in two remaining blocks they were asked to remember the digits or their position in the grid and to recognize them after the letter recall. In the ignore condition the expected effect of P300 decrease with letter position was found, especially at posterior electrode sites. When subjects had to process the preceding digit grid there was no position effect of P300 caused by small P300 amplitudes in the ERPs that were elicited by the first letters of the sequence. This effect was not specific to the modality of the interfering with the letter task information from the digit grid. We conclude that the grid information occupies working memory resources that are not available for the event-related processing of consonants to recall.

Adult↗

Mutational analysis of the functional domains of the large subunit of the isozyme form of wheat initiation factor eIF4F.

The isozyme form of plant eukaryotic initiation factor 4F (eIF(iso)4F) contains two subunits: p28, a cap-binding protein, and p86. To identify the functional domains of p86, truncations of the p86 cDNA were made, and the protein was expressed in Escherichia coli and purified. The deletion mutants were tested for the ability to bind the p28 subunit by two methods. In addition, these deletion mutants were evaluated in vitro by the ability to catalyze eIF4A and RNA-dependent ATP hydrolysis and to support polypeptide synthesis. The loss of the ability to bind p28 occurs within the first 90 amino acids of the N terminus and abrogates the ability of p86 to participate in translation initiation and bind to eIF4A, but does not affect ATP hydrolysis. Up to 299 amino acid residues from the C terminus of p86 must be deleted before an effect is observed on the ATP hydrolysis activity. Thus, the p28 binding and ATP hydrolysis activities appear to lie on two separate domains and are functionally uncoupled. In addition, at least a portion of the eIF4A binding domain appears to be in close proximity to the p28 binding domain and is also uncoupled from the ATP hydrolysis activity.

Adenosine Triphosphate↗

Sequences of two expressed sequence tags (EST) from rice encoding different cap-binding proteins.

Wheat has been shown to have two forms of the cap-binding protein that participate in the initiation of translation. To identify cap-binding proteins from other higher plant species, the expressed sequence tag (EST) database was searched. Several rice ESTs were identified with similarity to both forms of the wheat cap-binding proteins. Two of the rice ESTs were obtained and the cDNA sequences completed. The deduced amino acid sequences of the rice cap-binding proteins are compared to the wheat cap-binding proteins and cap-binding proteins from Saccharomyces cerevisiae, Drosophila melanogaster, Xenopus laevis and human.

Amino Acid Sequence↗

Sequence of a cDNA encoding wheat eukaryotic protein synthesis initiation factor 4A.

A cDNA encoding wheat translation initiation factor 4A (eIF-4A) was isolated from a wheat cDNA library and sequenced. The deduced wheat eIF-4A amino acid sequence from the cDNA is compared to eIF-4A from Arabidopsis thaliana, tobacco, mouse and Saccharomyces cerevisiae. Putative RNA helicase motifs, and putative ATP-binding and hydrolysis sites are identified.

Amino Acid Sequence↗

The 5' and 3' untranslated regions of satellite tobacco necrosis virus RNA affect translational efficiency and dependence on a 5' cap structure.

Satellite tobacco necrosis virus RNA (STNV RNA) is a naturally uncapped viral RNA that contains 1239 nucleotides: 29 in the 5' untranslated region (UTR), 591 in the coding region and 619 in the 3' UTR. Mutations were made in the 5' and 3' UTRs, and the effects of these mutations on translational efficiency and cap independence were measured in an in vitro translation system from wheat germ. Removal of the first 12 nucleotides or 10 changes in the nucleotide sequence of the 5' UTR reduced translational efficiency approximately 3-fold; capping of these 5' mutant mRNAs restored their translational efficiencies. Truncation of the 3' UTR to nucleotide 627 or 700, or deletion of nucleotides 627-737, reduced translational efficiency more than 20-fold; capping of these 3' mutant mRNAs restored their translational efficiencies. These modifications in the 3' UTR increased the concentration of initiation factor 4F required for translation. Chimeric mRNAs were constructed which contained the coding region of rabbit alpha-globin mRNA and either the 5' UTR, 3' UTR, or both the 5' and 3' UTRs of STNV RNA. Both the 5' and 3' UTRs of STNV RNA were necessary to obtain cap-independent translation. These findings indicate that interaction between 5' UTR and the region between nucleotides 627 and 737 in the 3' UTR are required for cap-independent translation.

Animals↗

Isolation and sequence of the cDNAs encoding the subunits of the isozyme form of wheat protein synthesis initiation factor 4F.

The nucleotide sequences of the cDNAs for the two subunits, p82 and p28, of the isozyme form of wheat germ eukaryotic initiation factor 4F (eIF-(iso)4F) were determined. The cDNA for the p82 subunit encodes a polypeptide of 86,514 Da. The deduced amino acid sequence of p82 contains possible motifs for ATP binding, metal binding, and phosphorylation. The cDNA sequence for the small subunit, p28, which is a m7G cap-binding protein, encodes a polypeptide of 23,524 Da. The deduced amino acid sequence of p28 is similar (approximately 38%) to cap-binding proteins from yeast and mammals. The p28 of wheat eIF-(iso)4F does not contain a serine or threonine in the vicinity of the serine (Ser53) of mammalian cap-binding protein which is phosphorylated and shown to affect activity in mammalian cells.

Amino Acid Sequence↗

Sequence of a cDNA encoding the alpha-subunit of wheat translation elongation factor 1.

A cDNA encoding the alpha-subunit of wheat protein synthesis elongation factor 1 (EF-1 alpha) was isolated from a wheat cDNA expression library and sequenced. The deduced amino acid sequence is compared to EF-1 alpha from other species and to elongation factor Tu (EF-Tu) from Escherichia coli. Putative GTP-binding sites are identified.

Amino Acid Sequence↗

[The P300 component of the event-related brain potential in a short-term memory paradigm].

In numerous investigations the P300-component of the event-related-brain-potential (ERP) has proved a valid indicator of memory activities. The present study explores the amplitude of the P300 in an isolated short term memory task with variant difficulties. In two experiments the reproduction of senseless memory material was examined using the categories "syllables" and "consonants". One syllable was thereby synonymous with the so-called "chunk" which represents a subjective unit arising from grouping of memory material. The results show a significant increase in P300-amplitude in a line with memory load in both categories. On the contrary there is no significant difference in the amplitude of P300 between the two categories. These results suggest that the costs for storing one syllable are comparable with those for one consonant.

Adult↗

[The history of the concept stress].

The historical roots of the term are treated as well as the story of Selye's theory of biological stress as a nonspecific response of the body. In the discussion of preceding approaches to describe processes which today one would name stress, the importance of Francois Magendie and John Brown as precursors is accentuated. Additionally, neurophysiological and psychological aspects are discussed.

Animals↗

[Occupational medicine outpatient management of workers with responsibilities involving great psychological stress].

Some selected results from regular medical occupational fitness evaluations, the statistic of unfitness for work, the disablement of teachers and the occupational referred mortality were demonstrated in special consideration of workers with particular psychic load. From it conclusions were drawn for the development of methods for occupational medical fitness examinations and check ups, prophylactic and therapeutic programmes. The necessarity of scientific dealing with this problems was emphasized.

Adult↗