Biomedical subjects

U G Maier

Publications and source records attributed to U G Maier.

36 records · Page 2Linked to original sources

PubMed1994

The cryptomonad histone H4-encoding gene: structure and chromosomal localization.

Cryptomonads are unicellular flagellates whose plastids are surrounded by four membranes. A periplastidal compartment, containing eukaryote-type ribosomes, starch grains and a so-called nucleomorph, is located between the inner and outer membrane pairs. The nucleomorph has been shown to be the vestigial nucleus of a eukaryotic endosymbiont. In order to obtain more information about the chromatin structure of the nucleomorph and the host nuclear chromosomes, we studied the distribution of the histone, H4. H4 was not detectable in the nucleomorph by immunolocalization, thus supporting earlier findings by Gibbs [In: Wiesner et al. (Eds.), Experimental Phycology 1, 1990, pp. 145-157]. Likewise, no H4 DNA was demonstrable in the nucleomorph by Southern hybridization. Sequence analysis, and Southern and Northern blot data of a cryptomonad gene, H4, indicate an intermediate position for these genes between animals and plants.

Amino Acid Sequence↗

PubMed1994

The smallest known eukaryotic genomes encode a protein gene: towards an understanding of nucleomorph functions.

Cryptomonads are unicellular algae with plastids surrounded by four membranes. Between the two pairs of membranes lies a periplastidal compartment that harbours a DNA-containing organelle, termed the nucleomorph. The nucleomorph is the vestigial nucleus of a phototrophic, eukaryotic endosymbiont. Subcloning of parts of one nucleomorph chromosome revealed a gene coding for an Hsp70 protein. We demonstrate the expression of this nucleomorph protein-coding gene and present a model for protein transport from the host to the endosymbiont compartment.

Biological Transport↗

PubMed1994

Evidence that an amoeba acquired a chloroplast by retaining part of an engulfed eukaryotic alga.

Chlorarachniophytes are amoeboid algae with unusual chloroplasts. Instead of the usual two membranes that surround the chloroplasts of plants, green algae, and red algae, the chloroplasts of chlorarachniophytes have four bounding membranes. The extra membranes may reflect an unusual origin of chlorarachniophyte chloroplasts. Rather than inheriting the organelle directly from their ancestors, chlorarachniophytes may have adopted the chloroplast of an algal cell ingested as prey. Parts of the algal cell are postulated to remain within the amoeba as a reduced eukaryotic endosymbiont [Hibberd, D. J. & Norris, R. E. (1984) J. Phycol. 20, 310-330]. A small nucleus-like structure, proposed to be a vestige of the endosymbiont's nucleus, is located in a space between the second and third chloroplast membranes. We cloned and sequenced nuclear-type rRNA genes from chlorarachniophytes and found two highly divergent genes. In situ hybridization shows that one gene is expressed by the amoebal (host) nucleus and the other is expressed by the putative endosymbiont nucleus, suggesting that the latter is indeed a foreign genome. Transcripts from the endosymbiont gene accumulate in the small cytoplasmic compartment between the second and third chloroplast membranes, which we believe to be the remnant cytoplasm of the endosymbiont. Using the endosymbiont gene as a probe, we identified three small chromosomes belonging to the endosymbiont nucleus. This knowledge should allow a detailed molecular analysis of the role of the endosymbiont's genome and cytoplasm in the partnership.

Amoeba↗

PubMed1994

The SecY protein family: comparative analysis and phylogenetic relationships.

We have cloned and sequenced the plastome encoded secY homologue of the cryptomonad alga Pyrenomonas salina. In this study we have carried out a comparative analysis of all but one fully sequenced proteins of the SecY family. We present an alignment of 16 SecY family proteins, containing family signatures, putative transmembrane domains, consensus sequence, and conservation grade. A phylogenetic tree derived from the conserved blocks of the alignment reveals the relationships among this protein family. The tree shows division into two broad subfamilies, one consisting of prokaryotic and plastidal sequences and the other of eukaryotic as well as archaeal sequences.

Amino Acid Sequence↗

PubMed1994

Phylogenetic analysis of the stress-70 protein family.

The eukaryotic cyto-/nucleoplasmatic 70-kDa heat-shock protein (HSP70) has homologues in the endoplasmic reticulum as well as in bacteria, mitochondria, and plastids. We selected a representative subset from the large number of sequenced stress-70 family members which covers all known branches of the protein family and calculated and manually improved an alignment. Here we present the consensus sequence of the aligned proteins and putative nuclear localization signals (NLS) in the eukaryotic HSP70 homologues. The phylogenetic relationships of the stress-70 group family members were estimated by use of different computation methods. We present a phylogenetic tree containing all known stress-70 subfamilies and demonstrate the usefulness of stress-70 protein sequences for the estimation of intertaxonic phylogeny.

Amino Acid Sequence↗

PubMed1993

Phylogenetic relationships of HMG box DNA-binding domains.

HMG boxes were initially identified as DNA-binding domains of the human RNA polymerase I (pol I) transcription factor hUBF and the animal high-mobility-group (HMG) protein family HMG1. Since then, numerous sequences of HMG-box-containing HMG proteins and other DNA-binding proteins from several species have become available. By sequence comparisons of a selected range of HMG boxes from these proteins and the construction of phylogenetic trees we show that the HMG box is highly conserved between DNA-binding proteins of organisms from all three eukaryotic kingdoms and that HMG boxes are linked by distinct evolutionary relationships. In addition, most HMG boxes display comparable hydropathy profiles and amino acid arrangements, which could serve as nuclear targeting sequences.

Amino Acid Sequence↗

PubMed1992

Plastid DNA from Pyrenomonas salina (Cryptophyceae): physical map, genes, and evolutionary implications.

Cryptomonads are thought to have arisen from a symbiotic association between a eukaryotic flagellated host and a eukaryotic algal symbiont, presumably related to red algae. As organellar DNAs have proven to be useful tools in elucidating phylogenetic relationships, the plastid (pt) DNA of the cryptomonad alga Pyrenomonas salina has been characterized in some detail. A restriction map of the circular 127 kb ptDNA from Pyrenomonas salina was established. An inverted repeat (IR) region of about 5 kb separates two single-copy regions of 15 and 102 kb, respectively. It contains the genes for the small and large subunit of rRNA. Ten protein genes, coding for the large subunit of ribulose-1,5-bisphosphate carboxylase, the 47 kDa, 43 kDa and 32 kDa proteins of photosystem II, the ribosomal proteins L2, S7 and S11, the elongation factor Tu, as well as the alpha- and beta-subunits of ATP synthase, have been localized on the restriction map either by hybridization of heterologous gene probes or by sequence homologies. The gene for the plastidal small subunit (SSUr) RNA has been sequenced and compared to homologous SSU regions from the cyanobacterium Anacystis nidulans and plastids from rhodophytes, chromophytes, euglenoids, chlorophytes, and land plants. A phylogenetic tree constructed with the neighborliness method and indicating a relationship of cryptomonad plastids with those of red algae is presented.

Base Sequence↗

PubMed1992

Evolutionary analysis of the plastid-encoded gene for the alpha subunit of the DNA-dependent RNA polymerase of Pyrenomonas salina (Cryptophyceae).

The nucleotide sequence of the gene coding for the plastid-encoded alpha subunit of DNA-dependent RNA polymerase from the cryptomonad alga Pyrenomonas salina was determined. The deduced amino-acid sequence, corresponding to a 35.2 kDa polypeptide, was compared to homologues from other organisms. Evolutionary relationships were analyzed in detail by the parsimony method together with bootstrap analysis. The deduced phylogenetic tree shows that the cryptomonad gene is the most ancient type of known plastid-encoded RNA polymerase.

Amino Acid Sequence↗

PubMed1992

The four genomes of the alga Pyrenomonas salina (Cryptophyta).

Cryptomonads are a group of unicellular eukaryotic algae with unusual features. First, their plastids are surrounded by four membranes and second, between the two pairs of membranes there is a plasmatic compartment. This supernumerary eukaryotic compartment of the cryptomonad cell is devoid of mitochondria but contains starch grains, 80S ribosomes and a small vestigial eukaryotic nucleus called the nucleomorph. Isolation and characterization of the four genomes (from mitochondrion, plastid, nucleus and nucleomorph) of one cryptomonad, Pyrenomonas salina, demonstrates that the cryptomonads have originated from an unicellular organism related to green algae which endosymbiotically took up a eukaryotic protist related to the red algae.

Biological Evolution↗

PubMed1991

A eukaryotic genome of 660 kb: electrophoretic karyotype of nucleomorph and cell nucleus of the cryptomonad alga, Pyrenomonas salina.

Cryptomonads are unicellular algae with chloroplasts surrounded by four membranes. Between the inner and the outer pairs of membranes is a narrow plasmatic compartment which contains a nucleus-like organelle called the nucleomorph. Using pulsed field gel electrophoresis it is shown that the nucleomorph of the cryptomonad Pyrenomonas salina contains three linear chromosomes of 195 kb, 225 kb and 240 kb all of which encode rRNAs. Thus, this vestigial nucleus has a haploid genome size of 660 kb, harboring the smallest eukaryotic genome known so far. From the cell nucleus of P. salina at least 20 chromosomes ranging from 230 kb to 3.000 kb were fractionated. Here, the rDNA was detected on a single chromosome of about 2.500 kb.

Blotting, Southern↗

PubMed1991

Demonstration of nucleomorph-encoded eukaryotic small subunit ribosomal RNA in cryptomonads.

In cryptomonads, unicellular phototrophic flagellates, the plastid(s) is (are) located in a special narrow compartment which is bordered by two membranes; it harbours neither mitochondria nor Golgi dictyosomes but comprises eukaryotic ribosomes and starch grains together with a small organelle called the nucleomorph. The nucleomorph contains DNA and is surrounded by a double membrane with pores. It is thought to be the vestigial nucleus of a phototrophic eukaryotic endosymbiont. Cryptomonads are therefore supposed to represent an intermediate state in the evolution of complex plastids from endosymbionts. We have succeeded in isolating pure nucleomorph fractions, and can thus provide, using pulsed field gel electrophoresis, polymerase chain reaction and sequence analysis, definitive proof for the eukaryotic nature of the symbiont and its phylogenetic origin.

Animals↗

PubMed1990

Maize high mobility group proteins bind to CCAAT and TATA boxes of a zein gene promoter.

A 216-base pair promoter fragment of the 19-kDa zein storage protein gene pMS1, containing the CCAAT and TATA boxes, was analyzed for its in vitro interactions with purified high mobility group (HMG) proteins from maize endosperm tissue. It was found that the HMG proteins display a preferential binding of A/T-rich DNA regions like their animal counterparts, although the A/T content seems not to be the only determinant for binding specificity. Surprisingly, a specific binding of the HMG proteins occurs at the CCAAT and the TATA boxes as indicated by footprinting experiments.

Base Sequence↗

PubMed1990

Multiple proteins bind to the P2 promoter region of the zein gene pMS1 of maize.

A 216 bp promoter fragment of the 19 kDa protein zein gene pMS1, containing the CCAAT and TATA boxes, was analysed by a variety of techniques for in vitro interactions with nuclear proteins from endosperm tissue. HMG proteins were found to form stable complexes with these A/T-rich promoter sequences and several specific DNA-binding proteins appear to be involved in the formation of DNA-protein complexes with this fragment. A 29 bp region spanning the two CCAAT boxes was protected from DNase I digestion in footprinting experiments.

Base Sequence↗

PubMed1989

A nuclear casein type II kinase from maize endosperm phosphorylating HMG proteins.

A casein kinase of the type II was isolated and enriched from nuclear lysates of maize endosperm tissue. The kinase activity requires 10 mM Mg2+ for maximal activity, can utilize either ATP or GTP as phosphate donors and is inhibited by polyamines, heparin and monovalent cations. A substrate specificity of the kinase activity towards specific nuclear proteins is indicated by its phosphorylation of high mobility group (HMG) proteins isolated from endosperm and its lack of accepting histones as protein substrates.

Animals↗

PubMed1989

Specific binding of nuclear proteins to the promoter region of a maize nuclear rRNA gene unit.

The external spacer region of a nuclear rRNA gene unit of maize was analyzed for binding of nuclear proteins and revealed binding to the promoter region and to the 200-base pair repeat elements upstream of the promoter. The promoter binding site was further characterized by DNase I and exonuclease III foot-printing as well as by gel shift and South-Western experiments, revealing the binding of several proteins. Competition experiments indicate that the protein which interacts with the repeat units also binds to the promoter region. A model for the combined action of both binding regions in the regulation of transcription is presented.

Binding, Competitive↗

PubMed1987

Binding of a nuclear factor to a consensus sequence in the 5' flanking region of zein genes from maize.

The genomic organization of the zein structural genes and of regulatory loci influencing their expression suggests that control of zein gene expression will involve interactions between cis elements in the flanking DNA sequences and products from trans-acting genes. The interaction between fragments from the 5' flanking region of a zein gene and specific, double-stranded oligonucleotides with crude nuclear extracts from maize endosperm have been studied by nitrocellulose filter binding, gel retention and DNase I footprinting assays. Specific binding of a nuclear factor was observed and the exact position of the protein binding site was determined. The 22-nt binding site included 14 bp of a 15-bp sequence conserved in all zein genes.

Consensus Sequence↗

PubMedDate not supplied

cDNA cloning of a Sec61 homologue from the cryptomonad alga Pyrenomonas salina.

Sec61 is an endoplasmic reticulum transmembrane protein involved in the process of translocation of proteins across this membrane. To-date, the only cloned genes for Sec61 are derived from mammals and yeast. In this paper, we present the first full-length cDNA from a sec61 gene of a plant cell. Comparison of the predicted protein sequence with all known Sec61 proteins, as well as with the bacterial/plastome-encoded homologue SecY, demonstrates a high degree of similarity among the SecY/Sec61 family.

Amino Acid Sequence↗

PubMedDate not supplied

The presence of a nucleomorph hsp70 gene is a common feature of Cryptophyta and Chlorarachniophyta.

Cryptomonad algae and Chlorarachniophyta are evolutionary chimaeras derived from the engulfment of an eukaryotic phototrophic endosymbiont by a eukaryotic host cell. Although much reduced, the endosymbiont's eukaryotic plasmatic compartment still contains a nucleus, the so-called nucleomorph. These nucleomorphs carry the smallest known eukaryotic genomes. We have characterized the genomes of several cryptomonads and a Chlorarachnion species by means of PFGE (pulsed-field gel electrophoresis). Hybridization studies with small subunit rDNA were used to identify the nucleomorph chromosomes. We also performed hybridization experiments with an hsp70 probe to estimate the distribution of this gene among the different algal species. The evolutionary, genetical, and physiological implications of our studies are discussed. A model on the possible function of the nucleomorph hsp70 gene products is presented.

Blotting, Southern↗