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R Maleszka

Publications and source records attributed to R Maleszka.

At least 19 recordsLinked to original sources
PubMed2000

Analysis of Drosophila yellow-B cDNA reveals a new family of proteins related to the royal jelly proteins in the honeybee and to an orphan protein in an unusual bacterium Deinococcus radiodurans.

The yellow locus in Drosophila is involved in both cuticle development and behaviour. However, the function of the encoded protein is unknown. Here we have characterised the sequence and expression pattern of a new Drosophila gene, designated yellow-B, encoding a 453-amino-acid protein that is 57% identical to Yellow. High levels of yellow-B mRNA are present in the larval-pupal stages, but the gene is also expressed in the head. Bioinformatics analysis indicates that the Drosophila genome encodes at least 7 members of the Yellow family distributed among chromosomes 2, 3, and X. The Yellow proteins are related to the Royal Jelly proteins and have no relatives in other non-insect metazoan species. Interestingly, a Yellow-like protein is encoded by the genome of a radiation tolerant bacterium, Deinococcus radiodurans.

Amino Acid Sequence↗
PubMed2000

Molecular cloning and expression analysis of a cDNA encoding a glutamate transporter in the honeybee brain.

We have cloned and characterized a cDNA encoding a putative glutamate transporter, Am-EAAT, from the brain of the honeybee, Apis mellifera. The 543-amino-acid AmEAAT gene product shares the highest sequence identity (54%) with the human EAAT2 subtype. Am-EAAT is expressed predominantly in the brain, and its transcripts are abundant in the optic lobes and inner compact Kenyon cells of the mushroom bodies (MBs), with most other regions of the brain showing lower levels of Am-EAAT expression. High levels of Am-EAAT message are found in pupal stages, possibly indicating a role for glutamate in the developing brain.

ATP-Binding Cassette Transporters↗
PubMed2000

Pharmacological interference with glutamate re-uptake impairs long-term memory in the honeybee, apis mellifera.

The role of glutamate in the central nervous system of invertebrates is poorly understood. In the present study we examined the effects of a glutamate transporter inhibitor, L-trans-2,4-pyrrolidine dicarboxylate (L-trans-2,4-PDC), on memory formation in the honeybee following a three-trial classical conditioning of the proboscis extension reflex (PER). Pre-training injections of the drug have no effect on acquisition and short-term (1 h) memory, but impair long-term (24 h), associative olfactory memory in a dose-dependent manner. This effect is transient and the amnesiac individuals can be re-trained successfully 48 h after injections. Our results suggest that glutamatergic neurons in the honeybee brain, in particular those found in the mushroom bodies (MBs), may be part of the circuitry involved in processing of long-term olfactory memory. Such a role for this neurotransmitter is consistent with our previous results showing that glutamate and glutamate transporter(s) are localised in regions of the honeybee brain implicated in higher order processing.

Amnesia↗
PubMed2000

Deus ex genomix.

Explore the source record for details and available documents.

Animals↗
PubMed1998

Arginine kinase is highly expressed in the compound eye of the honey bee, Apis mellifera.

We have cloned and sequenced a 1.68-kb cDNA encoding arginine kinase in the honey bee, Apis mellifera. The predicted protein shows a high level of identity to known arginine kinases in invertebrates and to other proteins belonging to the conserved family of ATP: guanidino phospho-transferases. The pattern of expression of arginine kinase has been investigated for the first time in various tissues including the brain, antennae and compound eye. Our results show that three isoforms of arginine kinase, transcribed from a single gene, are expressed in a characteristic pattern in major tissues of the honey bee. Arginine kinase mRNA is relatively abundant in the central nervous system and in the antennae. However, the highest level of expression, that is at least two to three times higher than in the brain, is found in the compound eye of the bee. By contrast, the levels of mRNAs encoding another metabolically important enzyme, alpha-glycerolphosphate dehydrogenase (alpha-GPDH), are low in the eye. These findings suggest that arginine kinase is an important component of the energy releasing mechanism in the visual system that has high and fluctuating energy demands. Furthermore, our results support the role of phosphagen kinases in energy transport in polarised cells and are consistent with the role of arginine kinase as an energy shuttle that delivers ATP generated by mitochondria to high energy-requiring processes, such as massive membrane turnover and pigment regeneration in the retina.

Amino Acid Sequence↗
PubMed1998

Data transferability from model organisms to human beings: insights from the functional genomics of the flightless region of Drosophila.

At what biological levels are data from single-celled organisms akin to a Rosetta stone for multicellular ones? To examine this question, we characterized a saturation-mutagenized 67-kb region of the Drosophila genome by gene deletions, transgenic rescues, phenotypic dissections, genomic and cDNA sequencing, bio-informatic analysis, reverse transcription-PCR studies, and evolutionary comparisons. Data analysis using cDNA/genomic DNA alignments and bio-informatic algorithms revealed 12 different predicted proteins, most of which are absent from bacterial databases, half of which are absent from Saccharomyces cerevisiae, and nearly all of which have relatives in Caenorhabditis elegans and Homo sapiens. Gene order is not evolutionarily conserved; the closest relatives of these genes are scattered throughout the yeast, nematode, and human genomes. Most gene expression is pleiotropic, and deletion studies reveal that a morphological phenotype is seldom observed when these genes are removed from the genome. These data pinpoint some general bottlenecks in functional genomics, and they reveal the acute emerging difficulties with data transferability above the levels of genes and proteins, especially with complex human phenotypes. At these higher levels the Rosetta stone analogy has almost no applicability. However, newer transgenic technologies in Drosophila and Mus, combined with coherency pattern analyses of gene networks, and synthetic neural modeling, offer insights into organismal function. We conclude that industrially scaled robogenomics in model organisms will have great impact if it can be realistically linked to epigenetic analyses of human variation and to phenotypic analyses of human diseases in different genetic backgrounds.

Animals↗
PubMed1997

The dodo gene family encodes a novel protein involved in signal transduction and protein folding.

Recent studies in yeast, Drosophila and humans have revealed the existence of a highly conserved gene encoding a novel protein, Dodo, comprised of four modules: a WW domain, involved in protein-protein interactions, a peptidyl-prolyl cis-trans isomerase (PPIase) domain belonging to a recently described third family of PPIases involved in protein folding and unfolding, a nuclear localization motif and finally, a long, surface-exposed alpha-helix that is likely to be involved in binding to a cell cycle serine/threonine kinase. The genetic, molecular, biochemical and structural data are reviewed in the context of the potential biological properties of this new protein family.

Amino Acid Sequence↗
PubMed1997

Molecular cloning, by a novel approach, of a cDNA encoding a putative olfactory protein in the labial palps of the moth Cactoblastis cactorum.

We have used the CapFinder technology, without the library construction step, to amplify and clone full-length cDNAs expressed in the labial palps (CO2-sensing organs) of the moth Cactoblastis cactorum. The validity of our approach is exemplified by the sequence analysis of a 597-bp cDNA clone, designated CLP-1, that contains a 390-bp open reading frame (ORF) flanked by motifs characteristic to a full-length cDNA. The ORF in CLP-1 encodes a predicted polypeptide that is 47% identical to a novel protein, OS-D, found exclusively in the olfactory antennal segment of Drosophila melanogaster. Both CLP-1 and OS-D have primary structures that do not bear sequence similarity to any previously characterised proteins including odorant-binding proteins (OBPs) in vertebrates and pheromone-binding proteins (PBPs) in moths. Although they share features common to OBPs and PBPs, such as the presence of signal peptides and cysteine motifs, they clearly belong to a distinct class of olfactory proteins that appear to be unique to insects. The relative abundance of the CLP-1 message in the labial palps of females leads to the suggestion that this protein is involved in the CO2-sensing cascade. Our results suggest that the experimental procedure can be used as an alternative, rapid method to identify genes expressed in a particular organ, or tissue, especially in situations when the amount of available tissue is a limiting factor.

Amino Acid Sequence↗
PubMed1997

An essential cell division gene of Drosophila, absent from Saccharomyces, encodes an unusual protein with tubulin-like and myosin-like peptide motifs.

Null mutations at the misato locus of Drosophila melanogaster are associated with irregular chromosomal segregation at cell division. The consequences for morphogenesis are that mutant larvae are almost devoid of imaginal disk tissue, have a reduction in brain size, and die before the late third-instar larval stage. To analyze these findings, we isolated cDNAs in and around the misato locus, mapped the breakpoints of chromosomal deficiencies, determined which transcript corresponded to the misato gene, rescued the cell division defects in transgenic organisms, and sequenced the genomic DNA. Database searches revealed that misato codes for a novel protein, the N-terminal half of which contains a mixture of peptide motifs found in alpha-, beta-, and gamma-tubulins, as well as a motif related to part of the myosin heavy chain proteins. The sequence characteristics of misato indicate either that it arose from an ancestral tubulin-like gene, different parts of which underwent convergent evolution to resemble motifs in the conventional tubulins, or that it arose by the capture of motifs from different tubulin genes. The Saccharomyces cerevisiae genome lacks a true homolog of the misato gene, and this finding highlights the emerging problem of assigning functional attributes to orphan genes that occur only in some evolutionary lineages.

Amino Acid Sequence↗
PubMed1996

The Drosophila melanogaster dodo (dod) gene, conserved in humans, is functionally interchangeable with the ESS1 cell division gene of Saccharomyces cerevisiae.

We have sequenced the region of DNA adjacent to and including the flightless (fli) gene of Drosophila melanogaster and molecularly characterized four transcription units within it, which we have named tweety (twe), flightless (fli), dodo (dod), and penguin (pen). We have performed deletion and transgenic analysis to determine the consequences of the quadruple gene removal. Only the flightless gene is vital to the organism; the simultaneous absence of the other three allows the overriding majority of individuals to develop to adulthood and to fly normally. These gene deletion results are evaluated in the context of the redundancy and degeneracy inherent in many genetic networks. Our cDNA analyses and data-base searches reveal that the predicted dodo protein has homologs in other eukaryotes and that it is made up of two different domains. The first, designated WW, is involved in protein-protein interactions and is found in functionally diverse proteins including human dystrophin. The second is involved in accelerating protein folding and unfolding and is found in Escherichia coli in a new family of peptidylprolyl cis-trans isomerases (PPIases; EC 5.2.1.8). In eukaryotes, PPIases occur in the nucleus and the cytoplasm and can form stable associations with transcription factors, receptors, and kinases. Given this particular combination of domains, the dodo protein may well participate in a multisubunit complex involved in the folding and activation of signaling molecules. When we expressed the dodo gene product in Saccharomyces cerevisiae, it rescued the lethal phenotype of the ESS1 cell division gene.

Amino Acid Sequence↗
PubMed1994

A gene homologous to that encoding UDP galactose-4-epimerase is inducible by xylose in the yeast Pachysolen tannophilus.

Using DNA sequencing, we have identified a fragment of genomic DNA from Pachysolen tannophilus that is homologous to the GAL10 gene in yeasts and galE gene in bacteria, both encoding the UDP galactose-4-epimerase (EC 5.1.3.2). This gene (designated PtGAL10) is equally efficiently inducible by D-galactose (a hexose) and D-xylose (a pentose). The encoded protein shows the highest similarity to the homologous Kluyveromyces lactis protein. This includes the N-terminal domain that is not present in homologous polypeptides from other organisms.

Amino Acid Sequence↗
PubMed1994

The in vivo effects of ethidium bromide on mitochondrial and ribosomal DNA in Candida parapsilosis.

The ability of Candida parapsilosis to grow in the presence of high levels of ethidium bromide (EB) has been explored to study the effects of this intercalating dye on DNA in vivo. By employing confocal microscopy we have determined that EB penetrates the cellular membranes and binds rapidly to the nucleolus, whereas mitochondrial DNA becomes stained after a longer exposure to this dye. No detectable staining of the nucleus has been detected under these conditions. Electrophoretic studies of both undigested and restricted DNAs confirm that the nuclear DNA is unaffected by high levels of EB, with the exception of the rDNA-bearing chromosome that undergoes significant structural alterations in the presence of EB. Moreover, the hybridization signal with the rDNA probe is proportionally reduced in samples obtained from cultures grown in the presence of EB, suggesting that the average copy number of rRNA genes in these cultures may be affected. In striking contrast to other fungal species, the linear organelle genome in C. parapsilosis retains its structural and functional integrity in the presence of high concentrations of EB.

Candida↗
PubMed1993

Yeasts have a four-fold variation in ribosomal DNA copy number.

By employing pulsed-field gel electrophoresis we have determined the size of the rDNA cluster in wild-type yeast strains representing genera of Candida, Kluyveromyces, Pachysolen, Schizosaccharomyces and Torulaspora. Although the genome size of the examined species is similar (12.3-13.9 Mb), at least a four-fold variation has been observed between the lowest amount of rDNA repeats in P. tannophilus (28) and the highest in C. glabrata and S. poombe (> 115). In two species the rDNA cluster is represented by two loci, residing either in one (S. pombe) or two chromosomes (C. glabrata).

Candida↗
PubMed1993

Electrophoretic analysis of the nuclear and organellar genomes in the ultra-small alga Cyanidioschyzon merolae.

Electrophoretic analysis reveals that the nucleus of the ultra-small eukaryotic alga Cyanidioschyzon merolae contains approximately 11.7 x 10(6) base pairs (11.7 Mb) of DNA. This compact genome is fragmented into 15 small chromosomes ranging in size from 410 to 1700 kb. The migratory behaviour of chloroplast DNA is consistent with the presence of a circular plastid genome of about 170 kb. The conformation of mitochondrial DNA resembles that in yeasts and fungi and is predominantly linear and heterogenous in size.

Cell Nucleus↗
PubMed1993

Single-stranded regions in yeast mitochondrial DNA revealed by pulsed-field gel electrophoresis.

The Klenow fragment of E. coli DNA polymerase together with pulsed-field gel electrophoresis (PFGE) have been used to investigate the presence of single-stranded DNA (ssDNA) regions in yeast (Torulopsis glabrata) DNA. Electrophoretic profiles of total DNA from Rho+ (wild type) and Rho0 (no mitochondrial DNA) strains demonstrate that this method mediates the incorporation of labelled dATP into mitochondrial DNA (mtDNA), but not into chromosomal DNA. The majority of ssDNA (> 62%) has been found associated with the electrophoretically inert component, localized on the top of PFGE gels. Treatment with single-stranded nucleases allows the resolution of this immobile fraction into fast migrating, linear molecules of a heterogeneous size. The possibility that single-stranded tracts and their recombinogenic properties are responsible for the trapping of DNA in pulsed-field gels is discussed.

DNA Replication↗
PubMed1992

Electrophoretic profiles of mitochondrial plasmids in Neurospora suggest they replicate by a rolling circle mechanism.

Migratory behaviour of mitochondrial plasmids from Neurospora crassa Mauriceville-1c and N. intermediate LaBelle has been studied by pulsed field gel electrophoresis (PFGE). Electrophoretic profiles demonstrate that long, linear molecules of a heterogeneous size are the prevailing form of plasmid DNA in vivo. Circular forms represent less than 8-9% of plasmid DNA. Single stranded DNA regions are abundant and lead to electrophoretic inertia of a significant amount of plasmid DNA. These profiles indicate that both plasmids replicate by the recombination dependent rolling circle mechanism.

DNA Replication↗