scholarly journals Molecular Diversity of the Syndinean Genus Euduboscquella Based on Single-Cell PCR Analysis

2011 ◽  
Vol 78 (2) ◽  
pp. 334-345 ◽  
Author(s):  
Tsvetan R. Bachvaroff ◽  
Sunju Kim ◽  
Laure Guillou ◽  
Charles F. Delwiche ◽  
D. Wayne Coats
Keyword(s):  
Large Subunit ◽  
Small Subunit ◽  
Base Change ◽  
Pcr Analysis ◽  
Ssu Rrna ◽  
Compensatory Base Change ◽  
Content Type ◽  
Single Cell Pcr ◽  
Group I ◽  
Lsu Rrna

ABSTRACTThe genusEuduboscquellais one of a few described genera within the syndinean dinoflagellates, an enigmatic lineage with abundant diversity in marine environmental clone libraries based on small subunit (SSU) rRNA. The region composed of the SSU through to the partial large subunit (LSU) rRNA was determined from 40 individual tintinnid ciliate loricae infected withEuduboscquellasampled from eight surface water sites in the Northern Hemisphere, producing seven distinct SSU sequences. The corresponding host SSU rRNA region was also amplified from eight host species. The SSU tree ofEuduboscquellaand syndinean group I sequences from environmental clones had seven well-supported clades and one poorly supported clade across data sets from 57 to 692 total sequences. The genusEuduboscquellaconsistently formed a supported monophyletic clade within a single subclade of group I sequences. For most parasites with identical SSU sequences, the more variable internal transcribed spacer (ITS) to LSU rRNA regions were polymorphic at 3 to 10 sites. However, inE. cachonithere was variation between ITS to LSU copies at up to 20 sites within an individual, while in a parasite ofTintinnopsisspp., variation between different individuals ranged up to 19 polymorphic sites. However, applying the compensatory base change model to the ITS2 sequences suggested no compensatory changes within or between individuals with the same SSU sequence, while one to four compensatory changes between individuals with similar but not identical SSU sequences were found. Comparisons between host and parasite phylogenies do not suggest a simple pattern of host or parasite specificity.

scholarly journals Pre-AIDS Era Isolates of Pneumocystis carinii f. sp. hominis: High Genotypic Similarity with Contemporary Isolates

1998 ◽  
Vol 36 (1) ◽  
pp. 90-93 ◽  
Author(s):  
Anthony G. Tsolaki ◽  
Pieter Beckers ◽  
Ann E. Wakefield
Keyword(s):  
Large Subunit ◽  
Pneumocystis Carinii ◽  
Small Subunit ◽  
Its Sequence ◽  
Small Subunit Rrna ◽  
Aids Pandemic ◽  
Genes Encoding ◽  
Lsu Rrna ◽  
Large Subunit Rrna ◽  
Sequence Types

Isolates of Pneumocystis carinii f. sp.hominis were examined from six individuals who died ofP. carinii pneumonia between 1968 and 1981 and who had underlying immunodeficiencies which were not due to human immunodeficiency virus infection. DNA sequence variation was analyzed in the genes encoding the mitochondrial large subunit rRNA (mt LSU rRNA), the internal transcribed spacer (ITS) regions of the nuclear rRNA, the arom locus, and the mitochondrial small subunit rRNA. No major variations were observed when these isolates were compared to isolates from HIV-infected individuals. A small number of minor differences were detected. A new position at which variation occurred in the mt LSU rRNA was observed in one sample. Three new ITS sequence types were identified. A total of nine different ITS sequence types were found in the six samples. Mixed infection with different ITS sequence types of P. carinii f. sp. hominis was observed in four of the six samples. The ITS locus was the most informative of the four loci for distinguishing among the isolates ofP. carinii f. sp. hominis. The data suggest that isolates of P. carinii f. sp. hominis from before the AIDS pandemic are genetically very similar to those currently found in HIV-infected individuals.

Ficophagus chaozhouensis n. sp. (Nematoda: Aphelenchoididae), an associate of Ficus hirta Vahl var. roxburghii (Miq.) King in China

Nematology ◽  
2020 ◽  
Vol 22 (3) ◽  
pp. 299-312
Author(s):  
Wensheng Zeng ◽  
Dayuan Zhang ◽  
Jianghua Huang ◽  
Yongsan Zeng ◽  
Weimin Ye ◽  
...  
Keyword(s):  
Large Subunit ◽  
Nematode Species ◽  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Monophyletic Clade ◽  
Male Tail ◽  
Lsu Rrna Gene ◽  
Ficus Hirta ◽  
Tail Tip

Summary A new nematode species was recovered from the syconia of Ficus hirta var. roxburghii from Chaozhou, Guangdong, China. It is described herein as Ficophagus chaozhouensis n. sp. and is characterised by possessing the combined characters of a short post-uterine sac, excretory pore located near the head, amoeboid sperm, three pairs of subventral papillae on the male tail, rounded male tail tip with mucron (occasionally swollen), absence of gubernaculum (or apophysis), a blunt rosethorn-shaped spicule without a terminal cucullus, and a digitate rostrum with a broadly squared tip. Ficophagus chaozhouensis n. sp. was separated from other sequenced species by differences in the partial small subunit (SSU) rRNA gene and D2-D3 expansion segments of the large subunit (LSU) rRNA gene. Phylogenetic analysis with LSU D2-D3 expansion segment sequences suggested that F. chaozhouensis n. sp. is clustered in the same highly supported monophyletic clade with F. aculeata, F. maxima and F. yoponensis, and is sister to F. aculeata.

scholarly journals Ciliate Environmental Diversity Can Be Underestimated by the V4 Region of SSU rDNA: Insights from Species Delimitation and Multilocus Phylogeny of Pseudokeronopsis (Ciliophora, Spirotrichea)

2019 ◽  
Vol 7 (11) ◽  
pp. 493 ◽  
Author(s):  
Zhan ◽  
Li ◽  
Xu
Keyword(s):  
Species Delimitation ◽  
High Throughput Sequencing ◽  
Sequence Similarity ◽  
Large Subunit ◽  
Small Subunit ◽  
Base Change ◽  
Published Data ◽  
Environmental Diversity ◽  
5.8S Rdna ◽  
Eukaryotic Microorganisms

Metabarcoding and high-throughput sequencing methods have greatly improved our understanding of protist diversity. Although the V4 region of small subunit ribosomal DNA (SSU-V4 rDNA) is the most widely used marker in DNA metabarcoding of eukaryotic microorganisms, doubts have recently been raised about its suitability. Here, using the widely distributed ciliate genus Pseudokeronopsis as an example, we assessed the potential of SSU-V4 rDNA and four other nuclear and mitochondrial markers for species delimitation and phylogenetic reconstruction. Our studies revealed that SSU-V4 rDNA is too conservative to distinguish species, and a threshold of 97% and 99% sequence similarity detected only one and three OTUs, respectively, from seven species. On the basis of the comparative analysis of the present and previously published data, we proposed the multilocus marker including the nuclear 5.8S rDNA combining the internal transcribed spacer regions (ITS1-5.8S-ITS2) and the hypervariable D2 region of large subunit rDNA (LSU-D2) as an ideal barcode rather than the mitochondrial cytochrome c oxidase subunit 1 gene, and the ITS1-5.8S-ITS2 as a candidate metabarcoding marker for ciliates. Furthermore, the compensating base change and tree-based criteria of ITS2 and LSU-D2 were useful in complementing the DNA barcoding and metabarcoding methods by giving second structure and phylogenetic evidence.

scholarly journals Community Structure Analysis of Methanogens Associated with Rumen Protozoa Reveals Bias in Universal Archaeal Primers

2012 ◽  
Vol 78 (11) ◽  
pp. 4051-4056 ◽  
Author(s):  
Lisa D. Tymensen ◽  
Tim A. McAllister
Keyword(s):  
Community Structure ◽  
Species Composition ◽  
Structure Analysis ◽  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Content Type ◽  
Ssu Rrna Gene ◽  
Rumen Protozoa ◽  
Primer Sets

ABSTRACTThe diversity of protozoan-associated methanogens in cattle was investigated using five universal archaeal small-subunit (SSU) rRNA gene primer sets.Methanobrevibacterspp. and rumen cluster C (distantly related toThermoplasmaspp.) were predominant. Significant differences in species composition among libraries indicate that some primers used previously to characterize rumen methanogens exhibit biased amplification.

scholarly journals Coordination of Synthesis and Assembly of a Modular Membrane-Associated [NiFe]-Hydrogenase Is Determined by Cleavage of the C-Terminal Peptide

2015 ◽  
Vol 197 (18) ◽  
pp. 2989-2998 ◽  
Author(s):  
Claudia Thomas ◽  
Enrico Muhr ◽  
R. Gary Sawers
Keyword(s):  
Signal Peptide ◽  
Large Subunit ◽  
Small Subunit ◽  
Catalytic Subunit ◽  
Regulatory Function ◽  
Accessory Protein ◽  
Membrane Association ◽  
Content Type ◽  
Hydrogenase 2

ABSTRACTDuring biosynthesis of [NiFe]-hydrogenase 2 (Hyd-2) ofEscherichia coli, a 15-amino-acid C-terminal peptide is cleaved from the catalytic large subunit precursor, pro-HybC. This peptide is removed only after NiFe(CN)2CO cofactor insertion by the Hyp accessory protein machinery has been completed, suggesting that it has a regulatory function during enzyme maturation. We show here that inhypmutants that fail to synthesize and insert the NiFe cofactor, and therefore retain the peptide, the Tat (twin-arginine translocon) signal peptide on the small subunit HybO is not removed and the subunit is degraded. In a mutant lacking the large subunit, the Tat signal peptide was also not removed from pre-HybO, indicating that the mature large subunit must actively engage the small subunit to elicit Tat transport. We validated the proposed regulatory role of the C-terminal peptide in controlling enzyme assembly by genetically removing it from the precursor of HybC, which allowed assembly and Tat-dependent membrane association of a HybC-HybO heterodimer lacking the NiFe(CN)2CO cofactor. Finally, genetic transfer of the C-terminal peptide from pro-HyaB, the large subunit of Hyd-1, onto HybC did not influence its dependence on the accessory protein HybG, a HypC paralog, or the specific protease HybD. This indicates that the C-terminal peptideper seis not required for interaction with the Hyp machinery but rather suggests a role of the peptide in maintaining a conformation of the protein suitable for cofactor insertion. Together, our results demonstrate that the C-terminal peptide on the catalytic subunit controls biosynthesis, assembly, and membrane association of Hyd-2.IMPORTANCE[NiFe]-hydrogenases are multisubunit enzymes with a catalytic subunit containing a NiFe(CN)2CO cofactor. Results of previous studies suggested that after synthesis and insertion of the cofactor by the Hyp accessory proteins, this large subunit changes conformation upon proteolytic removal of a short peptide from its C terminus. We show that removal of this peptide is necessary to allow the cleavage of the Tat signal peptide from the small subunit with concomitant membrane association of the heterodimer to occur. Genetic removal of the C-terminal peptide from the large subunit allowed productive interaction with the small subunit and Tat-dependent membrane insertion of a NiFe cofactor-free enzyme. Results based on swapping of C-terminal peptides between hydrogenases suggest that this peptide governs enzyme assembly via a conformational switch.

scholarly journals Identification and Characterization of the Rhizobium sp. Strain GIN611 Glycoside Oxidoreductase Resulting in the Deglycosylation of Ginsenosides

2011 ◽  
Vol 78 (1) ◽  
pp. 242-249 ◽  
Author(s):  
Eun-Mi Kim ◽  
Juhan Kim ◽  
Joo-Hyun Seo ◽  
Jun-Seong Park ◽  
Duck-Hee Kim ◽  
...  
Keyword(s):  
Enrichment Culture ◽  
Large Subunit ◽  
Small Subunit ◽  
Compound K ◽  
Oxidation Activity ◽  
Content Type ◽  
Molecular Masses ◽  
Ginsenoside Compound K ◽  
Identification And Characterization

ABSTRACTUsing enrichment culture,Rhizobiumsp. strain GIN611 was isolated as having activity for deglycosylation of a ginsenoside, compound K (CK). The purified heterodimeric protein complex fromRhizobiumsp. GIN611 consisted of two subunits with molecular masses of 63.5 kDa and 17.5 kDa. In the genome, the coding sequence for the small subunit was located right after the sequence for the large subunit, with one nucleotide overlapping. The large subunit showed CK oxidation activity, and the deglycosylation of compound K was performed via oxidation of ginsenoside glucose by glycoside oxidoreductase. Coexpression of the small subunit helped soluble expression of the large subunit in recombinantEscherichia coli. The purified large subunit also showed oxidation activity against other ginsenoside compounds, such as Rb1, Rb2, Rb3, Rc, F2, CK, Rh2, Re, F1, and the isoflavone daidzin, but at a much lower rate. When oxidized CK was extracted and incubated in phosphate buffer with or without enzyme, (S)-protopanaxadiol [PPD(S)] was detected in both cases, which suggests that deglycosylation of oxidized glucose is spontaneous.

scholarly journals Molybdenum-Containing Nicotine Hydroxylase Genes in a Nicotine Degradation Pathway That Is a Variant of the Pyridine and Pyrrolidine Pathways

2015 ◽  
Vol 81 (24) ◽  
pp. 8330-8338 ◽  
Author(s):  
Hao Yu ◽  
Hongzhi Tang ◽  
Yangyang Li ◽  
Ping Xu
Keyword(s):  
Inductively Coupled Plasma ◽  
Large Subunit ◽  
Sole Source ◽  
Small Subunit ◽  
Binding Domain ◽  
Mass Spectrometry Analysis ◽  
Resting Cells ◽  
Content Type ◽  
Spectrometry Analysis ◽  
Nicotine Degradation

ABSTRACTOchrobactrumsp. strain SJY1 utilizes nicotine as a sole source of carbon, nitrogen, and energy via a variant of the pyridine and pyrrolidine pathways (the VPP pathway). Several strains and genes involved in the VPP pathway have recently been reported; however, the first catalyzing step for enzymatic turnover of nicotine is still unclear. In this study, a nicotine hydroxylase for the initial hydroxylation step of nicotine degradation was identified and characterized. The nicotine hydroxylase (VppA), which converts nicotine to 6-hydroxynicotine in the strain SJY1, is encoded by two open reading frames (vppASandvppAL[subunits S and L, respectively]). ThevppAgenes were heterologously expressed in the non-nicotine-degrading strainsEscherichia coliDH5α andPseudomonas putidaKT2440; only thePseudomonasstrain acquired the ability to degrade nicotine. The small subunit of VppA contained a [2Fe-2S] cluster-binding domain, and the large subunit of VppA contained a molybdenum cofactor-binding domain; however, an FAD-binding domain was not found in VppA. Resting cells cultivated in a molybdenum-deficient medium had low nicotine transformation activity, and excess molybdenum was detected in the purified VppA by inductively coupled plasma-mass spectrometry analysis. Thus, it is demonstrated that VppA is a two-component molybdenum-containing hydroxylase.

scholarly journals A Novel Spirooxindole Derivative Inhibits the Growth of Leishmania donovani Parasites bothIn VitroandIn Vivoby Targeting Type IB Topoisomerase

2016 ◽  
Vol 60 (10) ◽  
pp. 6281-6293 ◽  
Author(s):  
Sourav Saha ◽  
Chiranjit Acharya ◽  
Uttam Pal ◽  
Somenath Roy Chowdhury ◽  
Kahini Sarkar ◽  
...  
Keyword(s):  
Leishmania Donovani ◽  
Nuclear Dna ◽  
Large Subunit ◽  
Peritoneal Macrophages ◽  
Small Subunit ◽  
Drug Resistant ◽  
Wild Type ◽  
Content Type ◽  
Fluorescence Studies ◽  
Topoisomerase Ib

ABSTRACTVisceral leishmaniasis is a fatal parasitic disease, and there is an emergent need for development of effective drugs against this neglected tropical disease. We report here the development of a novel spirooxindole derivative,N-benzyl-2,2′α-3,3′,5′,6′,7′,7α,α′-octahydro-2methoxycarbonyl-spiro[indole-3,3′-pyrrolizidine]-2-one (compound 4c), which inhibitsLeishmania donovanitopoisomerase IB (LdTopIB) and kills the wild type as well as drug-resistant parasite strains. This compound inhibits catalytic activity of LdTopIB in a competitive manner. Unlike camptothecin (CPT), the compound does not stabilize the DNA-topoisomerase IB cleavage complex; rather, it hinders drug-DNA-enzyme covalent complex formation. Fluorescence studies show that the stoichiometry of this compound binding to LdTopIB is 2:1 (mole/mole), with a dissociation constant of 6.65 μM. Molecular docking with LdTopIB using the stereoisomers of compound 4c produced two probable hits for the binding site, one in the small subunit and the other in the hinge region of the large subunit of LdTopIB. This spirooxindole is highly cytotoxic to promastigotes ofL. donovaniand also induces apoptosis-like cell death in the parasite. Treatment with compound 4c causes depolarization of mitochondrial membrane potential, formation of reactive oxygen species inside parasites, and ultimately fragmentation of nuclear DNA. Compound 4c also effectively clears amastigote forms of wild-type and drug-resistant parasites from infected mouse peritoneal macrophages but has less of an effect on host macrophages. Moreover, compound 4c showed strong antileishmanial efficacies in the BALB/c mouse model of leishmaniasis. This compound potentially can be used as a lead for developing excellent antileishmanial agents against emerging drug-resistant strains of the parasite.

scholarly journals Microbes in High Arctic Snow and Implications for the Cold Biosphere

2011 ◽  
Vol 77 (10) ◽  
pp. 3234-3243 ◽  
Author(s):  
Tommy Harding ◽  
Anne D. Jungblut ◽  
Connie Lovejoy ◽  
Warwick F. Vincent
Keyword(s):  
Microbial Mats ◽  
High Arctic ◽  
Small Subunit ◽  
Transport Processes ◽  
The Arctic ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Content Type ◽  
Ssu Rrna Gene ◽  
Aerial Transport

ABSTRACTWe applied molecular, microscopic, and culture techniques to characterize the microbial communities in snow and air at remote sites in the Canadian High Arctic (Ward Hunt Island, Ellesmere Island, and Cornwallis Island, latitudes 74 to 83oN). Members of theBacteriaandEukaryawere prevalent in the snow, and their small subunit (SSU) rRNA gene signatures indicated strong local aerial transport within the region over the preceding 8 months of winter snowpack accumulation. Many of the operational taxonomic units (OTUs) were similar to previously reported SSU rRNA gene sequences from the Arctic Ocean, suggesting the importance of local aerial transport processes for marine microbiota. More than 47% of the cyanobacterial OTUs in the snow have been previously found in microbial mats in the region, indicating that this group was also substantially derived from local sources. Viable cyanobacteria isolated from the snow indicated free exchange between the snow and adjacent mat communities. Other sequences were most similar to those found outside the Canadian Arctic but were from snow, lake and sea ice, glaciers and permafrost, alpine regions, Antarctica, and other regions of the Arctic, supporting the concept of global distribution of microbial ecotypes throughout the cold biosphere.

scholarly journals Clb6-Cdc28 Promotes Ribonucleotide Reductase Subcellular Redistribution during S Phase

2017 ◽  
Vol 38 (6) ◽  
Author(s):  
Xiaorong Wu ◽  
Xiuxiang An ◽  
Caiguo Zhang ◽  
Mingxia Huang
Keyword(s):  
Dna Damage ◽  
Ribonucleotide Reductase ◽  
Large Subunit ◽  
Small Subunit ◽  
S Phase ◽  
Nuclear Retention ◽  
Content Type ◽  
Dntp Pool ◽  
Checkpoint Kinases ◽  
Enhanced Sensitivity

ABSTRACTA tightly controlled cellular deoxyribonucleotide (deoxynucleoside triphosphate [dNTP]) pool is critical for maintenance of genome integrity. One mode of dNTP pool regulation is through subcellular localization of ribonucleotide reductase (RNR), the enzyme that catalyzes the rate-limiting step of dNTP biosynthesis. InSaccharomyces cerevisiae, the RNR small subunit, Rnr2-Rnr4, is localized to the nucleus, whereas the large subunit, Rnr1, is cytoplasmic. As cells enter S phase or encounter DNA damage, Rnr2-Rnr4 relocalizes to the cytoplasm to form an active holoenzyme complex with Rnr1. Although the DNA damage-induced relocalization requires the checkpoint kinases Mec1-Rad53-Dun1, the S-phase-specific redistribution does not. Here, we report that the S-phase cyclin–cyclin-dependent kinase (CDK) complex Clb6-Cdc28 controls Rnr2-Rnr4 relocalization in S phase. Rnr2 contains a consensus CDK site and exhibits Clb6-dependent phosphorylation in S phase. Deletion ofCLB6or removal of the CDK site results in an increased association of Rnr2 with its nuclear anchor Wtm1, nuclear retention of Rnr2-Rnr4, and an enhanced sensitivity to the RNR inhibitor hydroxyurea. Thus, we propose that Rnr2-Rnr4 redistribution in S phase is triggered by Clb6-Cdc28-mediated phosphorylation of Rnr2, which disrupts the Rnr2-Wtm1 interaction and promotes the release of Rnr2-Rnr4 from the nucleus.

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