Unequal contribution of two paralogous CENH3 variants in cowpea centromere function

AbstractIn most diploids the centromere-specific histone H3 (CENH3), the assembly site of active centromeres, is encoded by a single copy gene. Persistance of two CENH3 paralogs in diploids species raises the possibility of subfunctionalization. Here we analysed both CENH3 genes of the diploid dryland crop cowpea. Phylogenetic analysis suggests that gene duplication of CENH3 occurred independently during the speciation of Vigna unguiculata. Both functional CENH3 variants are transcribed, and the corresponding proteins are intermingled in subdomains of different types of centromere sequences in a tissue-specific manner together with the kinetochore protein CENPC. CENH3.2 is removed from the generative cell of mature pollen, while CENH3.1 persists. CRISPR/Cas9-based inactivation of CENH3.1 resulted in delayed vegetative growth and sterility, indicating that this variant is needed for plant development and reproduction. By contrast, CENH3.2 knockout individuals did not show obvious defects during vegetative and reproductive development. Hence, CENH3.2 of cowpea is likely at an early stage of pseudogenization and less likely undergoing subfunctionalization.

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Unequal contribution of two paralogous centromeric histones to function the cowpea centromere

10.1101/2020.01.07.897074 ◽

2020 ◽

Cited By ~ 1

Author(s):

Takayoshi Ishii ◽

Martina Juranić ◽

Shamoni Maheshwari ◽

Fernanda de Oliveira Bustamante ◽

Maximilian Moritz Vogt ◽

...

Keyword(s):

Genome Duplication ◽

Early Stage ◽

Generative Cell ◽

Reproductive Development ◽

Duplication Event ◽

Genome Duplication Event ◽

A Genome ◽

Specific Manner ◽

Different Types ◽

Drought And Heat Stress

AbstractThe legume cowpea (Vigna unguiculata, 2n=2x=22) has significant tolerance to drought and heat stress. Here we analysed and manipulated cowpea centromere-specific histone H3 (CENH3) genes, aiming to establish a centromere-based doubled-haploid method for use in genetic improvement of this dryland crop in future. Cowpea encodes two functional CENH3 variants (CENH3.1 and CENH3.2) and two CENH3 pseudogenes. Phylogenetic analysis suggests that gene duplication of CENH3 occurred independently during the speciation of V. unguiculata and the related V. mungo without a genome duplication event. Both functional cowpea CENH3 variants are transcribed, and the corresponding proteins are intermingled in subdomains of different types of centromere sequences in a tissue-specific manner together with the outer kinetochore protein CENPC. CENH3.2 is removed from the generative cell of mature pollen, while CENH3.1 persists. Differences between both CENH3 paralogs are restricted to the N-terminus. The complete CRISPR/Cas9-based inactivation of CENH3.1 resulted in delayed vegetative growth and sterility, indicating that this variant is needed for plant development and reproduction. By contrast, CENH3.2 knockout individuals did not show obvious defects during vegetative and reproductive development, suggesting that the gene is an early stage of subfunctionalization or pseudogenization.One-sentence summaryThe two paralogous centromeric histones (CENH3) of cowpea contribute unequal to the function of the centromere.

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Identification of a single-copy gene encoding a Type I chlorophyll a/b-binding polypeptide of photosystem I in Arabidopsis thaliana

Physiologia Plantarum ◽

10.1034/j.1399-3054.1992.840410.x ◽

1992 ◽

Vol 84 (4) ◽

pp. 561-567 ◽

Cited By ~ 1

Author(s):

Poul E. Jensen ◽

Michael Kristensen ◽

Tine Hoff ◽

Jan Lehmbeck ◽

Bjarne M. Stummann ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Chlorophyll A ◽

Photosystem I ◽

Single Copy ◽

Type I ◽

Single Copy Gene ◽

Gene Encoding ◽

Copy Gene

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A Mouse Single-Copy Gene,Gtf2i,the Homolog of HumanGTF2I,That Is Duplicated in the Williams–Beuren Syndrome Deletion Region

Genomics ◽

10.1006/geno.1997.5182 ◽

1998 ◽

Vol 48 (2) ◽

pp. 163-170 ◽

Cited By ~ 28

Author(s):

Yu-Ker Wang ◽

Luis A. Pérez-Jurado ◽

Uta Francke

Keyword(s):

Single Copy ◽

Single Copy Gene ◽

Deletion Region ◽

Copy Gene

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Real-Time PCR for Molecular Detection of Zoonotic and Non-Zoonotic Giardia spp. in Wild Rodents

Microorganisms ◽

10.3390/microorganisms9081610 ◽

2021 ◽

Vol 9 (8) ◽

pp. 1610

Author(s):

Christian Klotz ◽

Elke Radam ◽

Sebastian Rausch ◽

Petra Gosten-Heinrich ◽

Toni Aebischer

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Gastrointestinal Disease ◽

Single Copy ◽

Marker Genes ◽

Small Rodent ◽

Analytical Sensitivity ◽

Single Copy Gene ◽

Wild Rodents ◽

Copy Gene

Giardiasis in humans is a gastrointestinal disease transmitted by the potentially zoonotic Giardia duodenalis genotypes (assemblages) A and B. Small wild rodents such as mice and voles are discussed as potential reservoirs for G. duodenalis but are predominantly populated by the two rodent species Giardia microti and Giardia muris. Currently, the detection of zoonotic and non-zoonotic Giardia species and genotypes in these animals relies on cumbersome PCR and sequencing approaches of genetic marker genes. This hampers the risk assessment of potential zoonotic Giardia transmissions by these animals. Here, we provide a workflow based on newly developed real-time PCR schemes targeting the small ribosomal RNA multi-copy gene locus to distinguish G. muris, G. microti and G. duodenalis infections. For the identification of potentially zoonotic G. duodenalis assemblage types A and B, an established protocol targeting the single-copy gene 4E1-HP was used. The assays were specific for the distinct Giardia species or genotypes and revealed an analytical sensitivity of approximately one or below genome equivalent for the multi-copy gene and of about 10 genome equivalents for the single-copy gene. Retesting a biobank of small rodent samples confirmed the specificity. It further identified the underlying Giardia species in four out of 11 samples that could not be typed before by PCR and sequencing. The newly developed workflow has the potential to facilitate the detection of potentially zoonotic and non-zoonotic Giardia species in wild rodents.

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Tissue-specific expression of two gamma-glutamyl transpeptidase mRNAs with alternative 5' ends encoded by a single copy gene in the rat.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)39983-1 ◽

1990 ◽

Vol 265 (4) ◽

pp. 2352-2357

Author(s):

M N Chobert ◽

O Lahuna ◽

F Lebargy ◽

O Kurauchi ◽

M Darbouy ◽

...

Keyword(s):

Single Copy ◽

Single Copy Gene ◽

Gamma Glutamyl Transpeptidase ◽

Specific Expression ◽

Tissue Specific ◽

Gamma Glutamyl ◽

Tissue Specific Expression ◽

Copy Gene ◽

Glutamyl Transpeptidase

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A synthetic intron in a naturally intronless yeast pre-tRNA is spliced efficiently in vivo

Molecular and Cellular Biology ◽

10.1128/mcb.9.1.329-331.1989 ◽

1989 ◽

Vol 9 (1) ◽

pp. 329-331

Author(s):

M Winey ◽

I Edelman ◽

M R Culbertson

Keyword(s):

Saccharomyces Cerevisiae ◽

Genetic Analysis ◽

Single Copy ◽

Single Copy Gene ◽

Copy Gene

Saccharomyces cerevisiae glutamine tRNA(CAG) is encoded by an intronless, single-copy gene, SUP60. We have imposed a requirement for splicing in the biosynthesis of this tRNA by inserting a synthetic intron in the SUP60 gene. Genetic analysis demonstrated that the interrupted gene produces a functional, mature tRNA product in vivo.

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Novel chicken actin gene: third cytoplasmic isoform

Molecular and Cellular Biology ◽

10.1128/mcb.5.5.1151-1162.1985 ◽

1985 ◽

Vol 5 (5) ◽

pp. 1151-1162

Author(s):

D J Bergsma ◽

K S Chang ◽

R J Schwartz

Keyword(s):

Nucleotide Sequence ◽

Single Copy ◽

Actin Gene ◽

Single Copy Gene ◽

Gene Diversification ◽

Mrna Transcripts ◽

Copy Gene ◽

Actin Mrna ◽

Distinct Member ◽

Actin Protein

We identified a novel chicken actin gene. The actin protein deduced from its nucleotide sequence very closely resembles the vertebrate cytoplasmic actins; accordingly, we classified this gene as a nonmuscle type. We adopted the convention for indicating the nonmuscle actins of the class Amphibia (Vandekerckhove et al., J. Mol. Biol. 152:413-426) and denoted this gene as type 5. RNA blot analysis demonstrated that the type 5 actin mRNA transcripts accumulate in adult tissues in a pattern indicative of a nonmuscle actin gene. Genomic DNA blots indicated that the type 5 actin is a single copy gene and a distinct member of the chicken actin multigene family. Inspection of the nucleotide sequence revealed many features that distinguished the type 5 gene from all other vertebrate actin genes examined to date. These unique characteristics include: (i) an initiation Met codon preceding an Ala codon, a feature previously known only in plant actins, (ii) a single intron within the 5' untranslated region, with no interruptions in the coding portion of the gene, and (iii) an atypical Goldberg-Hogness box (ATAGAA) preceding the mRNA initiation terminus. These unusual features have interesting implications for actin gene diversification during evolution.

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