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M A Ragan

Publications and source records attributed to M A Ragan.

At least 37 records · Page 2Linked to original sources
PubMed1995

Characterization of the polyubiquitin gene in the marine red alga Gracilaria verrucosa.

We have cloned a nuclear gene (UBI6R) and corresponding cDNAs that encode polyubiquitin in the florideophycidean red alga Gracilaria verrucosa. The gene encodes a polyubiquitin composed of six tandem ubiquitin units, followed by a single glutamine residue. The deduced amino acid sequences are identical among all six units, and identical to the ubiquitin of the florideophyte Aglaothamnion neglectum. There is high sequence similarity among the red algal ubiquitins and those of animals, green plants, fungi and several protists. Only one polyubiquitin gene was found by Southern hybridization analysis of G. verrucosa nuclear DNA. The upstream region of the gene is rich in putative cis-acting transcription-regulatory elements, including a putative heat-responsive element. Poly(A) addition to UBI6R mRNA was observed in cDNAs at four different sites, implicating the sequences AATAAA and (or) AGTAAA as poly(A) addition signals. The polyubiquitin genes of red algae show features of concerted evolution, but appear to be subject to less sequence homogenization than those of animals.

Amino Acid Sequence↗
PubMed1995

Characterization of the nuclear gene encoding mitochondrial aconitase in the marine red alga Gracilaria verrucosa.

We have cloned a nuclear gene from the marine red alga Gracilaria verrucosa that encodes the complete 779 amino-acid mitochondrial aconitase (m-ACN), the first characterized from a photosynthetic organism. The N-terminal 28 deduced amino acids are predicted to constitute the mitochondrial transit peptide, the first described from a red alga. Putative transcriptional cis-acting elements were identified in the upstream untranslated region. The G. verrucosa m-ACN gene (m-ACN) is present in a single copy and is located ca. 1.5 kb upstream from the single-copy polyubiquitin gene. The single spliceosomal intron is located near the 5' end of the region encoding the mature m-ACN in precisely the same location and phase as intron 2 in Caenorhabditis elegans m-ACN; sequences at its 3' and 5' splice junctions and at the predicted lariat branch point conform well to the eukaryote consensus sequences. Multiple protein-sequence alignment of m-ACN, bacterial aconitase (b-ACN) and iron-responsive element-binding protein (IRE-BP), and phylogenetic analyses, revealed that m-ACN does not share a recent common ancestry with either b-ACN or IRE-BP.

Aconitate Hydratase↗
PubMed1995

Cloning and characterization of the nuclear gene and cDNAs for triosephosphate isomerase of the marine red alga Gracilaria verrucosa.

cDNAs and an intronless single-copy nuclear gene (TPI1) encoding triosephosphate isomerase have been cloned and sequenced from the marine red alga Gracilaria verrucosa. The predicted amino-acid sequence of TPI1 is readily alignable with those of other known TPIs; 26 of 27 active-site residues and 19 of 26 intersubunit-contact residues are identical between TPIs of G. verrucosa and/or animals and green plants. A partial cDNA sequence of a second TPI gene (TPI2), presumably encoding plastid-localized TPI, was recovered by PCR and demonstrated by phylogenetic analysis to be red algal; no TP12 cDNA or genomic clones could be recovered. Genomic Southern analysis demonstrated that at least two TPI-like genes are present in the nuclear DNA of G. verrucosa.

Amino Acid Sequence↗
PubMed1995

The nuclear gene and cDNAs encoding cytosolic glyceraldehyde-3-phosphate dehydrogenase from the marine red alga Gracilaria verrucosa: cloning, characterization and phylogenetic analysis.

We have cloned and sequenced the single-copy nuclear gene (GapC) encoding the complete 335-amino acid cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC) from the red alga Gracilaria verrucosa. The proline residue which contributes to the specificity of NAD+ binding in other GAPC-like proteins is present. Putative regulatory regions, including GC-rich regions, a GATA element, and 11-base T- and T/G-clusters, but excluding TATA- and CCAAT-boxes, were identified upstream. Two types of GapC cDNAs differing in polyadenylation site were characterized. An 80-bp phase-two spliceosomal intron was identified in a novel position interrupting the highly conserved cofactor-coding region I. The G. verrucosa GAPC was easily aligned with other known GAPC-type sequences. Inferred phylogenetic trees place red algae among the eukaryote crown taxa, although with modest bootstrap support and without stable resolution among related GAPC lineages.

Amino Acid Sequence↗
PubMed1994

A molecular phylogeny of the marine red algae (Rhodophyta) based on the nuclear small-subunit rRNA gene.

A phylogeny of marine Rhodophyta has been inferred by a number of methods from nucleotide sequences of nuclear genes encoding small subunit rRNA from 39 species in 15 orders. Sequence divergences are relatively large, especially among bangiophytes and even among congeners in this group. Subclass Bangiophycidae appears polyphyletic, encompassing at least three lineages, with Porphyridiales distributed between two of these. Subclass Florideophycidae is monophyletic, with Hildenbrandiales, Corallinales, Ahnfeltiales, and a close association of Nemaliales, Acrochaetiales, and Palmariales forming the four deepest branches. Cermiales may represent a convergence of vegetative and reproductive morphologies, as family Ceramiaceae is at best weakly related to the rest of the order, and one of its members appears to be allied to Gelidiales. Except for Gigartinales, for which more data are required, the other florideophyte orders appear distinct and taxonomically justified. A good correlation was observed with taxonomy based on pit-plug ultrastructure. Tests under maximum-likelihood and parsimony of alternative phylogenies based on structure and chemistry refuted suggestions that Acrochaetiales is the most primitive florideophyte order and that Gelidiales and Hildenbrandiales are sister groups.

Base Sequence↗
PubMed1994

Cloning and characterization of the nuclear gene encoding plastid glyceraldehyde-3-phosphate dehydrogenase from the marine red alga Gracilaria verrucosa.

The single-copy nuclear gene (GapA), encoding the plastid-localized glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of the marine red alga Gracilaria verrucosa, has been cloned and sequenced. The GapA transcriptional initiation site was located 49 bp upstream of the start codon, and a putative TATA box was found 54 bp farther upstream. A spliceosomal intron was identified in the transit-peptide-encoding region in a position very similar to intron 1 of GapA and GapB of higher plants; no introns occur in the region encoding the mature protein. These observations provisionally suggest that both red algae and higher plants descend from a single ancestral photosynthetic eukaryote, i.e. that a single endosymbiotic event gave rise to red algal and higher-plant plastids.

Amino Acid Sequence↗
PubMed1994

The 16S-like, 5.8S and 23S-like rRNAs of the two varieties of Cryptococcus neoformans: sequence, secondary structure, phylogenetic analysis and restriction fragment polymorphisms.

The nucleotide sequences of the 16S-like, 5.8S and 23S-like rDNAs from the two varieties of Cryptococcus neoformans, C. neoformans var. neoformans and C. neoformans var. gattii, were determined. The rRNA locus has the typical eukaryote organization of 16S-5.8S-23S with the 16S-like and 5.8S rRNA genes separated by a 124-nucleotide spacer and the 5.8S and 23S-like rRNA genes separated by a 187-nucleotide spacer in each strain. The C. neoformans var. neoformans and C. neoformans var. gattii 16S-like, 5.8S and 23S-like rRNAs are, respectively 1802, 158, and 3358 nucleotides in length and share > 99% nucleotide sequence identity, a finding which strongly supports the present taxonomic classification of two varieties within a species. Comparative structure analysis was used to construct secondary-structure models for the deduced 16S-like and 5.8S-23S-like rRNA sequences, which are similar to those of other fungal rRNAs. The C. neoformans 16S-like and 23S-like rRNA sequences were aligned with other eukaryote sequences based on secondary and higher-order structures predicted by comparative structure analysis for phylogenetic analysis. There was good correspondence between the 16S-like and 23S-like derived phylogenetic trees. The closest known fungal relative is Trichosporon beigelii. Southern blot analysis revealed one C. neoformans strain with two types of DNA repeats coding for rRNA which differed in size by about 1000 bp. Restriction fragment length polymorphisms in the rDNA locus provide useful markers for the study of epidemiology and pathogenesis of C. neoformans infections.

Base Sequence↗
PubMed1994

Variant forms of a group I intron in nuclear small-subunit rRNA genes of the marine red alga Porphyra spiralis var. amplifolia.

A group IC1 intron occurs in nuclear small-subunit (18S) ribosomal RNA (SSU rRNA) genes of the marine red alga Porphyra spiralis var. amplifolia. This intron occurs at the same position as the self-splicing group IC1 introns in nuclear SSU rDNAs of the fungus Pneumocystis carinii and in the green alga Chlorella ellipsoidea and shares sequence identity with the Pneumocystis carinii intron in domains L1, P1, P2, and L2, outside the conserved core. Three size variants, differing in amount of sequence in L1, exist and are differentially distributed in geographically distinct populations. Preliminary data suggest that the largest variant can self-splice in vitro. Short open reading frames are present but do not correspond to known genes. Repeated nucleotide motifs, reminiscent of duplicated target sites of transposons or Alu elements, are associated with the intron and with one of the variant forms of L1. Insertions are present in nuclear SSU rDNAs of several other Porphyra species and of the red alga Bangia atropurpurea; insertionless rDNA variants also occur in several Porphyra species. Our observations are most readily explained by intron mobility, although it remains unclear how transfer could have been mediated between genomes of organisms as ecologically diverse as marine red algae, freshwater green algae, and a mammalian-pathogenic fungus.

Base Sequence↗
PubMed1994

Phylogenetic and molecular characterization of a 23S rRNA gene positions the genus Campylobacter in the epsilon subdivision of the Proteobacteria and shows that the presence of transcribed spacers is common in Campylobacter spp.

The nucleotide sequence of a 23S rRNA gene of Campylobacter coli VC167 was determined. The primary sequence of the C. coli 23S rRNA was deduced, and a secondary-structure model was constructed. Comparison with Escherichia coli 23S rRNA showed a major difference in the C. coli rRNA at approximately position 1170 (E. coli numbering) in the form of an extra sequence block approximately 147 bp long. PCR analysis of 31 other strains of C. coli and C. jejuni showed that 69% carried a transcribed spacer of either ca. 147 or ca. 37 bp. Comparison of all sequenced Campylobacter transcribed spacers showed that the Campylobacter inserts were related in sequence and percent G+C content. All Campylobacter strains carrying transcribed spacers in their 23S rRNA genes produced fragmented 23S rRNAs. Other strains which produced unfragmented 23S rRNAs did not appear to carry transcribed spacers at this position in their 23S rRNA genes. At the 1850 region (E. coli numbering), Campylobacter 23S rRNA displayed a base pairing signature most like that of the beta and gamma subdivisions of the class Proteobacteria, but in the 270 region, Campylobacter 23S rRNA displayed a helix signature which distinguished it from the alpha, beta, and gamma subdivisions. Phylogenetic analysis comparing C. coli VC167 23S rRNA and a C. jejuni TGH9011 (ATCC 43431) 23S rRNA with 53 other completely sequenced (eu)bacterial 23S rRNAs showed that the two campylobacters form a sister group to the alpha, beta, and gamma proteobacterial 23S rRNAs, a positioning consistent with the idea that the genus Campylobacter belongs to the epsilon subdivision of the class Proteobacteria.

Bacteria↗
PubMed1992

Matrix representation in reconstructing phylogenetic relationships among the eukaryotes.

The application of the method of matrix representation (MRP) addresses several problems associated with reconstructing phylogenetic relationships among composite organisms such as the eukaryotes. By allowing multiple molecular-sequence and non-molecular data sets to be combined into a single, relatively compact matrix for joint analysis, MRP addresses problems associated with the depth, diversification and specialization of eukaryotic lineages. Characters whose distributions may have been affected by endosymbiotic lateral transfer can be identified by compatibility analysis of MRP-generated hybrid matrices. In conjunction with variant-filtering techniques, MRP can be used to map patterns of lateral transfer in a given phylogenetic tree. These applications are illustrated with molecular-sequence and non-molecular data sets for eukaryotes.

Animals↗
PubMed1992

Phylogenetic inference based on matrix representation of trees.

Rooted phylogenetic trees can be represented as matrices in which the rows correspond to termini, and columns correspond to internal nodes (elements of the n-tree). Parsimony analysis of such a matrix will fully recover the topology of the original tree. The maximum size of the represented matrix depends only on the number of termini in the tree; for a tree derived from molecular sequences, the represented matrix may be orders of magnitude smaller than the original data matrix. Representations of multiple trees (which may or may not have identical termini) can readily be combined into a single matrix; columns of discrete-character-state data can be added and, if desired, weighted differentially. Parsimony analysis of the resulting composite matrix yields a hybrid supertree which typically provides greater resolution than conventional consensus trees. Use of this method is illustrated with examples involving multiple tRNA genes in organelles and multiple protein-coding genes in eukaryotes.

Animals↗
PubMed1988

Ribosomal RNA and the major lines of evolution: a perspective.

Does the "universal tree" based on small-subunit ribosomal RNA sequences show the phylogenetic relationship of all modern organisms? The answer is "yes" only if all these rRNAs are orthologous. Herein I argue that the major rRNA lineages (e.g. eubacterial, one or more archaebacterial and eukaryotic nucleocytoplasmic) probably arose from a divergent population of rRNAs in the progenote, antedating the universal common ancestral organism. Thus the major lineages of rRNA are probably not orthologous, but paralogous. The extrapolated date for the origin of the common ancestral small-subunit rRNA (3.6-4.7 x 10(9) years ago) is consistent with major rRNA lineages being paralogous. This perspective on the early evolution of genes and organisms rationalizes the presence of unexpected ribosomal characters in microsporidia, and bears on xenogenous and endogenous theories of the origin of the organelles in eukaryotes.

Biological Evolution↗
PubMed1979

Paired-ion reversed-phase high-performance liquid chromatography of phenol sulfates in synthetic mixtures, algal extracts and urine.

Phenol sulfate esters have been analyzed by paired-ion reversed-phase high-performance liquid chromatography. The method provided direct, rapid chromatography of phenol sulfates in crude extracts of the red alga (Polysiphonia lanosa (2,3-dibromo-4,5-dihydroxybenzyl alcohol 1',4-disulfate), of the brown alga Ascophyllum nodosum (1,2,3,5-tetrahydroxybenzene 2,5-disulfate), and in rat urine (resorcinol mono- and disulfates). Detector response (254 nm) was linear within the approximate range from 30--125 ng to 5--10 microgram. Semipreparative scale chromatography provided sufficient amounts of purified phenol sulfates for further analysis by paper electrophoresis.

Animals↗
PubMed1976

Physodes and the phenolic compounds of brown algae. Isolation and characterization of phloroglucinol polymers from Fucus vesiculosus (L.)

A series of vanillin-reactive compounds has been isolated from extracts of the brown alga Fucus vesiculosus. The first three members of this series have been examined by mass spectrometry, nuclear magnetic resonance spectrometry, and chemical methods, and were shown to be phloroglucinol, its phenyl-linked dimer, and a timer also composed of phloroglucinol units. Evidence is presented for the presence of tetrameric, pentameric, and hexameric phloroglucinol derivatives. Polymeric vanillin-reactive compounds were isolated and separated into two fractions. Oxidative hydrolysis of the major one produced phloroglucinol and related oligomers in low yields. The classical vanillin reactivity of physodes may thus be attributed to phloroglucinol-containing derivatives, as originally proposed by Crato (Crato, E.: Ber. Dtsch. Bot, Ges. 10, 295-302 (1892)).

Chromatography, Paper↗
PubMedDate not supplied

cDNA cloning and characterization of the nuclear gene encoding chloroplast glyceraldehyde-3-phosphate dehydrogenase from the marine red alga Gracilaria verrucosa.

Using a PCR-generated homologous probe, we have recovered a cDNA (GapA cDNA) encoding the complete 338 amino-acid chloroplast GAPDH of the marine red alga Gracilaria verrucosa, together with its 78 amino-acid transit peptide. This cDNA was readily aligned with chloroplast-localized GAPDH genes (GapA and GapB) of green plants. The proline residue which contributes to the specificity of NAD+ binding to cytosolic GAPDHs is absent from the deduced polypeptide chain of G. verrucosa GapA as is also the case in the chloroplast GAPDHs of plants. The transit peptide shows a high proportion of random coil, an amino-terminal Met-Ala dipeptide, a high content of hydroxylamino acids, and a net positive charge. The polyadenylation signal appears to be AGTAAA. Genomic Southern-hybridization data indicate that only one chloroplast-GAPDH gene may occur in G. verrucosa. Bootstrapped parsimony trees indicate that the G. verrucosa GapA gene is a sister group to plant chloroplast-GAPDH genes, and are most readily interpreted as showing that red algal and plant chloroplast-localized GAPDHs arose in a single endosymbiotic event.

Amino Acid Sequence↗