Cloning and sequencing of tubulin cDNAs from Artemia franciscana: evidence for differential expression of α- and β-tubulin genes

2009 ◽  
Vol 87 (6) ◽  
pp. 989-997 ◽  
Author(s):  
Katy A. Garant ◽  
Thomas H. MacRae
Keyword(s):  
Allelic Variation ◽  
Gene Families ◽  
Artemia Franciscana ◽  
Multiple Gene ◽  
Cdna Sequences ◽  
Life History Stages ◽  
Tubulin Genes ◽  
Tubulin Mrna ◽  
Developmental Phases ◽  
Β Tubulin

Tubulin is a heterodimeric protein composed of α- and β-tubulin. In most organisms, they are encoded by multiple gene families whose members are subject to differential regulation. The objective of the work described herein was to better understand tubulin gene expression in the extremophile Artemia franciscana To this end tubulin cDNAs were cloned and sequenced. αAT2, an α-tubulin cDNA, differed by one nucleotide from αAT1, a previously cloned Artemia cDNA. This change, possibly generated by allelic variation, caused an M313V substitution in α-tubulin. The amino acid sequence of β-tubulin encoded by βAT1, one of only a very limited number of cloned crustacean β-tubulin cDNA sequences yet available, and the first from Artemia, was similar to other β-tubulins. However, βAT1 possessed four degenerate TATA boxes in the 5′ untranslated region, although authentic TATA and CCAAT boxes occurred in the 3′ non-coding sequence. Analyses by quantitative PCR demonstrated that the amount of tubulin mRNA declined relative to total mRNA in progressive life history stages of Artemia and also that the organism contained more αAT2- than βAT1-tubulin mRNA at all developmental phases examined.

Evidence for translational control of β-tubulin synthesis during differentiation ofLeishmania donovani

Parasitology ◽  
1991 ◽  
Vol 103 (2) ◽  
pp. 197-205 ◽  
Author(s):  
M. Bhaumik ◽  
S. Das ◽  
S. Adhya
Keyword(s):  
Translational Control ◽  
Leishmania Donovani ◽  
Cell Types ◽  
Northern Blotting ◽  
Translational Efficiency ◽  
Rnase Protection ◽  
Tubulin Genes ◽  
Rabbit Reticulocyte Lysate System ◽  
Tubulin Mrna ◽  
Β Tubulin

Tubulin biosynthesis was rapidly induced during transformation of the mammalian (amastigote) stage of the kinetoplastid parasiteLeishmania donovanito flagellated promastigotes. However, transcription of β-tubulin genes occurred constitutively, as judged by nascent RNA synthesis in isolated nuclei and Northern blotting of steady-state mRNA. Two mRNA species of 2.2 and 2.4 kb were shared by the two cell-types, while a third 2.6 kb species, constituting about 20% of the total, was present in large amounts in promastigotes. RNase protection experiments demonstrated sequence micro-heterogeneity in the 5′-untranslated region, the pattern of which was identical in promastigotes and amastigotes. By primer extension assays, heterogeneity in the 5′-terminal cap structure of amastigote β-tubulin mRNA and differential pausing of reverse transcriptase within the mini-exon leader region were detected. These differences correlated with enhanced translational efficiency of tubulin mRNA from promastigotes in a rabbit reticulocyte lysate system. The results indicate that translational control plays a major role in tubulin induction duringL. donovanidifferentiation.

scholarly journals Characterization of the β-tubulin gene family in Ascaris lumbricoides and Ascaris suum and its implication for the molecular detection of benzimidazole resistance

2021 ◽  
Vol 15 (9) ◽  
pp. e0009777
Author(s):  
Sara Roose ◽  
Russell W. Avramenko ◽  
Stephen M. J. Pollo ◽  
James D. Wasmuth ◽  
Shaali Ame ◽  
...  
Keyword(s):  
Gene Family ◽  
Ascaris Lumbricoides ◽  
Ascaris Suum ◽  
Gene Families ◽  
Surveillance Systems ◽  
Nucleotide Polymorphisms ◽  
Drug Pressure ◽  
Tubulin Gene ◽  
Tubulin Genes ◽  
Β Tubulin

Background The treatment coverage of control programs providing benzimidazole (BZ) drugs to eliminate the morbidity caused by soil-transmitted helminths (STHs) is unprecedently high. This high drug pressure may result in the development of BZ resistance in STHs and so there is an urgent need for surveillance systems detecting molecular markers associated with BZ resistance. A critical prerequisite to develop such systems is an understanding of the gene family encoding β-tubulin proteins, the principal targets of BZ drugs. Methodology and principal findings First, the β-tubulin gene families of Ascaris lumbricoides and Ascaris suum were characterized through the analysis of published genomes. Second, RNA-seq and RT-PCR analyses on cDNA were applied to determine the transcription profiles of the different gene family members. The results revealed that Ascaris species have at least seven different β-tubulin genes of which two are highly expressed during the entire lifecycle. Third, deep amplicon sequencing was performed on these two genes in more than 200 adult A. lumbricoides (Ethiopia and Tanzania) and A. suum (Belgium) worms, to investigate the intra- and inter-species genetic diversity and the presence of single nucleotide polymorphisms (SNPs) that are associated with BZ resistance in other helminth species; F167Y (TTC>TAC or TTT>TAT), E198A (GAA>GCA or GAG>GCG), E198L (GAA>TTA) and F200Y (TTC>TAC or TTT>TAT). These particular SNPs were absent in the two investigated genes in all three Ascaris populations. Significance This study demonstrated the presence of at least seven β-tubulin genes in Ascaris worms. A new nomenclature was proposed and prioritization of genes for future BZ resistance research was discussed. This is the first comprehensive description of the β-tubulin gene family in Ascaris and provides a framework to investigate the prevalence and potential role of β-tubulin sequence polymorphisms in BZ resistance in a more systematic manner than previously possible.

scholarly journals Susceptible trichostrongyloid species mask presence of benzimidazole-resistant Haemonchus contortus in cattle

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Khalid M. Mohammedsalih ◽  
Jürgen Krücken ◽  
Ahmed Bashar ◽  
Fathel-Rahman Juma ◽  
Abdalhakaim A. H. Abdalmalaik ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Epidemiological Data ◽  
Nematode Species ◽  
Parasitic Nematodes ◽  
Molecular Approaches ◽  
Tubulin Genes ◽  
Faecal Egg Count ◽  
Third Stage ◽  
Before And After ◽  
Β Tubulin

Abstract Background Benzimidazole (BZ) anthelmintics are widely used to control infections with parasitic nematodes, but BZ resistance is an emerging threat among several nematode species infecting humans and animals. In Sudan, BZ-resistant Haemonchus contortus populations were recently reported in goats in South Darfur State. The objective of this study was to collect data regarding the situation of BZ resistance in cattle parasitic nematodes in South Darfur using phenotypic and molecular approaches, besides providing some epidemiological data on nematodes in cattle. Methods The faecal egg count reduction test and the egg hatch test (EHT) were used to evaluate benzimidazole efficacy in cattle nematodes in five South Darfur study areas: Beleil, Kass, Nyala, Rehed Al-Birdi and Tulus. Genomic DNA was extracted from pools of third-stage larvae (L3) (n = 40) during trials, before and after treatment, and pools of adult male Haemonchus spp. (n = 18) from abattoirs. The polymorphisms F167Y, E198A and F200Y in isotype 1 β-tubulin genes of H. contortus and H. placei were analysed using Sanger and pyrosequencing. Results Prevalence of gastro-intestinal helminths in cattle was 71% (313/443). Reduced albendazole faecal egg count reduction efficacy was detected in three study areas: Nyala (93.7%), Rehed Al-Birdi (89.7%) and Tulus (88.2%). In the EHT, EC50 values of these study areas ranged between 0.032 and 0.037 µg/ml thiabendazole. Genus-specific PCRs detected the genera Haemonchus, Trichostrongylus and Cooperia in L3 samples collected after albendazole treatment. Sanger sequencing followed by pyrosequencing assays did not detect elevated frequencies of known BZ resistance-associated alleles in codon F167Y, E198A and F200Y in isotype 1 β-tubulin gene of H. placei (≤ 11.38%). However, polymorphisms were detected in H. contortus and in samples with mixed infections with H. contortus and H. placei at codon 198, including E198L (16/58), E198V (2/58) and potentially E198Stop (1/58). All pooled L3 samples post-albendazole treatment (n = 13) were identified as H. contortus with an E198L substitution at codon 198. Conclusions To the knowledge of the authors, this is the first report of reduced albendazole efficacy in cattle in Sudan and is the first study describing an E198L substitution in phenotypically BZ-resistant nematodes collected from cattle.

scholarly journals Diversity of Neopestalotiopsis and Pestalotiopsis spp., Causal Agents of Guava Scab in Colombia

Plant Disease ◽  
2018 ◽  
Vol 102 (1) ◽  
pp. 49-59 ◽  
Author(s):  
Fernando Solarte ◽  
Carlos German Muñoz ◽  
Sajeewa S. N. Maharachchikumbura ◽  
Elizabeth Álvarez
Keyword(s):  
Central America ◽  
Internal Transcribed Spacer ◽  
Geographical Origin ◽  
Elongation Factor ◽  
Psidium Guajava ◽  
Economic Importance ◽  
Fruit Tree ◽  
Multiple Gene ◽  
Guava Leaves ◽  
Β Tubulin

Common guava (Psidium guajava L.) is a fruit tree of global economic importance. It is grown in Asia, South and Central America, and Hawaii for its exquisite aroma and flavor, and nutritional and medical properties. However, guava production is limited by guava scab, caused by fungi in the Pestalotiopsis genus. Characteristic symptoms of guava scab are corky, ovoid or round lesions on fruit surfaces. These lesions may thicken, affecting the flesh below and reducing fruit quality and commercial value. We characterized 81 isolates isolated from guava scab lesions on guava leaves and fruit in different regions of Colombia, and identified them as Pestalotiopsis and Neopestalotiopsis spp. We analyzed the morphology, pathogenicity, and genetic diversity of the isolates based on the sequences of the internal transcribed spacer, β-tubulin, and elongation factor genes. Isolates were morphologically, pathogenically, and genetically diverse but the diversity did not correlate with geographical origin, or guava cultivar or tissue from which the isolates were recovered. Selected monosporic isolates included in the multiple-gene analysis were identified as belonging to two genera: Neopestalotiopsis (65 isolates with versicolorous conidia) and Pestalotiopsis (4 isolates with concolorous conidia).

scholarly journals Genetic polymorphism of white mistletoe (Viscum album L.) in Ukraine

2021 ◽  
Vol 28 ◽  
pp. 36-41
Author(s):  
Yu. O. Bilonozhko ◽  
A. M. Rabokon ◽  
A. S. Postovoitova ◽  
L. O. Kalafat ◽  
N. S. Boiko ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Polyacrylamide Gel Electrophoresis ◽  
Viscum Album ◽  
Intron Length ◽  
Intron Length Polymorphism ◽  
Tubulin Genes ◽  
Silver Nitrate Staining ◽  
Dna Profiles ◽  
Different Parts ◽  
Β Tubulin

Aim. The aim of the study was to establish genetic differences between V. album growing in different parts of Ukraine. Methods. White mistletoe samples collected in different regions of Ukraine were used in the study. The method of estimating the intron length polymorphism of β-tubulin genes was used. Amplified DNA fragments were fractionated by non-denaturing polyacrylamide gel electrophoresis and visualized by silver nitrate staining. Results. The genotypes of 91 white mistletoe plants were analyzed. DNA profiles of white mistletoe with a specific amplicons of β-tubulin gene introns were obtained, which allowed to differentiate the samples from each other. Fingerprinting data were used for cluster analysis and dendrogram construction. Conclusions. It was found that the analyzed mistletoe samples did not differ by geographical factor and were characterized by a low level of genetic diversity in the studied samples. Keywords: Viscum album L., intron length polymorphism, β-tubulin, genetic variability, Ukraine.

scholarly journals Occurrence of a New Subclade of Leptosphaeria biglobosa in Western Australia

2008 ◽  
Vol 98 (3) ◽  
pp. 321-329 ◽  
Author(s):  
L. Vincenot ◽  
M. H. Balesdent ◽  
H. Li ◽  
M. J. Barbetti ◽  
K. Sivasithamparam ◽  
...  
Keyword(s):  
Oilseed Rape ◽  
Mating Type ◽  
Western Australia ◽  
Leptosphaeria Maculans ◽  
Stem Canker ◽  
Tubulin Genes ◽  
Wide Scale ◽  
Scale Characterization ◽  
Leptosphaeria Biglobosa ◽  
Β Tubulin

Stem canker of crucifers is caused by an ascomycete species complex comprising of two main species, Leptosphaeria maculans and L. biglobosa. These are composed of at least seven distinct subclades based on biochemical data or on sequences of internal transcribed spacer (ITS), the mating type MAT1-2 or fragments of actin or β-tubulin genes. In the course of a wide-scale characterization of the race structure of L. maculans from Western Australia, a few isolates from two locations failed to amplify specific sequences of L. maculans, i.e., the mating-type or minisatellite alleles. Based on both pathogenicity tests and ITS size, these isolates were classified as belonging to the L. biglobosa species. Parsimony and distance analyses performed on ITS, actin and β-tubulin sequences revealed that these isolates formed a new L. biglobosa subclade, more related to the Canadian L. biglobosa ‘canadensis’ subclade than to the L. biglobosa ‘australensis’ isolates previously described in Australia (Victoria). They are termed here as L. biglobosa ‘occiaustralensis’. These isolates were mainly recovered from resistant oilseed rape cultivars that included the Brassica rapa sp. sylvestris-derived resistance source, but not from the susceptible cv. Westar. The pathogenicity of L. biglobosa ‘occiaustralensis’ to cotyledons of most oilseed rape genotypes was higher than that of L. biglobosa ‘canadensis’ or L. biglobosa ‘australensis’ isolates.

Stimulation of tubulin gene transcription by deciliation of sea urchin embryos

1987 ◽  
Vol 7 (12) ◽  
pp. 4238-4246
Author(s):  
Z Y Gong ◽  
B P Brandhorst
Keyword(s):  
Gene Transcription ◽  
Sea Urchin ◽  
Elevated Level ◽  
Elevated Concentration ◽  
Tubulin Gene ◽  
Lytechinus Pictus ◽  
Tubulin Genes ◽  
Tubulin Mrna ◽  
Hypertonic Shock ◽  
Stimulation Of

Deciliation by hypertonic shock of embryos of the sea urchin Lytechinus pictus resulted in an increase in synthesis of alpha- and beta-tubulins, the consequence of an increased concentration of RNA encoding the tubulins. RNA run-on assays in isolated nuclei indicated that this response is due to a transient increase in the rate of synthesis of tubulin RNA beginning within 5 min of deciliation. This enhancement of tubulin gene transcription also occurred in deciliated embryos treated with the microtubule-depolymerizing agent colcemid; thus the reaction to deciliation is not a response to a reduction in concentration of unpolymerized tubulin utilized for ciliogenesis. In deciliated embryos treated with colcemid, the elevated level of tubulin RNA declined rapidly, due to its destabilization by the elevated concentration of unpolymerized tubulin. The increased transcription of tubulin genes is a response to the loss of cilia, not to the hypertonic shock, and occurs even when cilium regeneration is prevented. Inhibition of protein synthesis with puromycin or emetine did not prevent the transcriptional enhancement but stabilized tubulin mRNA, resulting in increased accumulation of tubulin mRNA after deciliation.

GENETICS OF THE TUBULIN GENE FAMILIES OF PHYSARUM

Genetics ◽  
1984 ◽  
Vol 108 (1) ◽  
pp. 143-164
Author(s):  
Tim Schedl ◽  
Judi Owens ◽  
William F Dove ◽  
Timothy G Burland
Keyword(s):  
Drug Resistance ◽  
Genetic Mapping ◽  
Gene Family ◽  
Gene Families ◽  
Resistance Locus ◽  
Tubulin Gene ◽  
Length Polymorphisms ◽  
Mapping Analysis ◽  
Mendelian Analysis ◽  
Β Tubulin

ABSTRACT The organization of the α- and β-tubulin gene families in Physarum was investigated by Mendelian analysis. Restriction endonuclease-generated DNA fragments homologous to α- and β-tubulin show length polymorphisms that can be used as markers for genetic mapping. Analysis of meiotic assortment among progeny of heterozygotes allowed α- and β-tubulin sequence loci to be defined. There are four unlinked α-tubulin sequence loci (altA, altB, altC and altD) and at least three unlinked β-tubulin sequence loci (betA, betB and betC). The α-tubulin loci are not linked to the β-tubulin loci. —Segregation of tubulin sequence loci with respect to ben mutations that confer resistance to antitubulin benzimidazole drugs was used to investigate whether any members of the α- or β-tubulin gene families are allelic to ben loci. The β-tubulin sequence locus betB is allelic to the resistance locus benD, the betA locus is probably allelic to benA and the α-tubulin sequence locus altC may be allelic to benC. The molecular implications of benzimidazole resistance phenotypes when only one of the expressed β-tubulin gene family members mutates to drug resistance are discussed in relation to tubulin function.

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