A novel tissue specific alternative splicing variant mitigates phenotypes in Ets2 frame-shift mutant models

AbstractE26 avian leukemia oncogene 2, 3′ domain (Ets2) has been implicated in various biological processes. An Ets2 mutant model (Ets2db1/db1), which lacks the DNA-binding domain, was previously reported to exhibit embryonic lethality caused by a trophoblast abnormality. This phenotype could be rescued by tetraploid complementation, resulting in pups with wavy hair and curly whiskers. Here, we generated new Ets2 mutant models with a frame-shift mutation in exon 8 using the CRISPR/Cas9 method. Homozygous mutants could not be obtained by natural mating as embryonic development stopped before E8.5, as previously reported. When we rescued them by tetraploid complementation, these mice did not exhibit wavy hair or curly whisker phenotypes. Our newly generated mice exhibited exon 8 skipping, which led to in-frame mutant mRNA expression in the skin and thymus but not in E7.5 Ets2em1/em1 embryos. This exon 8-skipped Ets2 mRNA was translated into protein, suggesting that this Ets2 mutant protein complemented the Ets2 function in the skin. Our data implies that novel splicing variants incidentally generated after genome editing may complicate the phenotypic analysis but may also give insight into the new mechanisms related to biological gene functions.

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BmCHSA-2b, a Lepidoptera specific alternative splicing variant of epidermal chitin synthase, is required for pupal wing development in Bombyx mori

Insect Biochemistry and Molecular Biology ◽

10.1016/j.ibmb.2017.06.017 ◽

2017 ◽

Vol 87 ◽

pp. 117-126 ◽

Cited By ~ 9

Author(s):

Guanfeng Xu ◽

Jie Zhang ◽

Hao Lyu ◽

Jia Liu ◽

Yang Ding ◽

...

Keyword(s):

Alternative Splicing ◽

Bombyx Mori ◽

Chitin Synthase ◽

Wing Development ◽

Splicing Variant ◽

Pupal Wing ◽

Specific Alternative ◽

Alternative Splicing Variant

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Ets2 frame-shift mutant models express in-frame mRNA by exon skipping that complements Ets2 function in the skin

10.1101/2020.09.27.316075 ◽

2020 ◽

Author(s):

Yuki Kishimoto ◽

Iori Nishiura ◽

Shunsuke Yuri ◽

Nami Yamamoto ◽

Masahito Ikawa ◽

...

Keyword(s):

Exon Skipping ◽

Cell Types ◽

Dna Binding Domain ◽

Biological Processes ◽

Tetraploid Complementation ◽

Embryonic Lethal ◽

Frame Shift ◽

Homozygous Mutant ◽

Summary Statement ◽

Ets Domain

AbstractThe Ets2 transcription factor has been implicated in various biological processes. An Ets2 mutant model, which lacks the DNA-binding domain (ETS domain), was previously reported to exhibit embryonic lethality caused by a trophoblast abnormality. This phenotype could be rescued by tetraploid complementation, resulting in pups with wavy hair.Here, we generated new Ets2 mutant models with deletions in exon 8 and with frame-shift mutations using the CRISPR/Cas9 method. Homozygous mutants could not be obtained by natural mating as previously reported. After rescuing with tetraploid complementation, homozygous mutant mice were generated, but these mice did not exhibit wavy hair phenotype. Our newly generated mice exhibited exon 8 skipping, which led to in-frame mutant mRNA expression in the skin and thymus but not in E7.5 embryos. As this in-frame mutation contained the ETS domain, the exon 8-skipped Ets2 mRNA was likely translated into protein in the skin that complemented the Ets2 function. Thus, these Ets2 mutant models, depending on the cell types, exhibited novel phenotypes due to exon skipping and are expected to be useful in several fields of research.Summary statementNew Ets2 mutant models showed embryonic lethal phenotype by a placental abnormality but did not exhibit a wavy hair phenotype as a previous model.

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An Insight into Oligopeptide Transporter 3 (OPT3) family proteins

Protein and Peptide Letters ◽

10.2174/0929866527666200625202028 ◽

2020 ◽

Vol 27 ◽

Author(s):

Fırat Kurt

Keyword(s):

Stress Responses ◽

Chelating Agent ◽

Biological Processes ◽

Biotic And Abiotic Stress ◽

Oligopeptide Transporter ◽

Binding Residues ◽

Fe Homeostasis ◽

Proteins Interactions ◽

Insight Into ◽

Selection Of

: Oligopeptide transporter 3 (OPT3) proteins are one of the subsets of OPT clade, yet little is known about these transporters. Therefore, homolog OPT3 proteins in several plant species were investigated and characterized using bioinformatical tools. Motif and co-expression analyses showed that OPT3 proteins may be involved in both biotic and abiotic stress responses as well as growth and developmental processes. AtOPT3 usually seemed to take part in Fe homeostasis whereas ZmOPT3 putatively interacted with proteins involved in various biological processes from plant defense system to stress responses. Glutathione (GSH), as a putative alternative chelating agent, was used in the AtOPT3 and ZmOPT3 docking analyses to identify their putative binding residues. The information given in this study will contribute to the understanding of OPT3 proteins’ interactions in various pathways and to the selection of potential ligands for OPT3s.

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Genetic and phenotypic analysis of the pathogenic potential of two novel Chlamydia gallinacea strains compared to Chlamydia psittaci

Scientific Reports ◽

10.1038/s41598-021-95966-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Marloes Heijne ◽

Martina Jelocnik ◽

Alexander Umanets ◽

Michael S. M. Brouwer ◽

Annemieke Dinkla ◽

...

Keyword(s):

Virulence Factors ◽

Opportunistic Pathogen ◽

Chlamydia Psittaci ◽

Phenotypic Analysis ◽

Pathogenic Potential ◽

The Family ◽

Chicken Eggs ◽

Sequence Types ◽

Insight Into ◽

Embryonated Chicken

AbstractChlamydia gallinacea is an obligate intracellular bacterium that has recently been added to the family of Chlamydiaceae. C. gallinacea is genetically diverse, widespread in poultry and a suspected cause of pneumonia in slaughterhouse workers. In poultry, C. gallinacea infections appear asymptomatic, but studies about the pathogenic potential are limited. In this study two novel sequence types of C. gallinacea were isolated from apparently healthy chickens. Both isolates (NL_G47 and NL_F725) were closely related to each other and have at least 99.5% DNA sequence identity to C. gallinacea Type strain 08-1274/3. To gain further insight into the pathogenic potential, infection experiments in embryonated chicken eggs and comparative genomics with Chlamydia psittaci were performed. C. psittaci is a ubiquitous zoonotic pathogen of birds and mammals, and infection in poultry can result in severe systemic illness. In experiments with embryonated chicken eggs, C. gallinacea induced mortality was observed, potentially strain dependent, but lower compared to C. psittaci induced mortality. Comparative analyses confirmed all currently available C. gallinacea genomes possess the hallmark genes coding for known and potential virulence factors as found in C. psittaci albeit to a reduced number of orthologues or paralogs. The presence of potential virulence factors and the observed mortality in embryonated eggs indicates C. gallinacea should rather be considered as an opportunistic pathogen than an innocuous commensal.

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Transcriptomic and phenotypic analysis of murine embryonic stem cell derived BMP2+ lineage cells: an insight into mesodermal patterning

Genome Biology ◽

10.1186/gb-2007-8-9-r184 ◽

2007 ◽

Vol 8 (9) ◽

pp. R184 ◽

Cited By ~ 19

Author(s):

Michael Doss ◽

Shuhua Chen ◽

Johannes Winkler ◽

Rita Hippler-Altenburg ◽

Margareta Odenthal ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Embryonic Stem ◽

Phenotypic Analysis ◽

Murine Embryonic Stem Cell ◽

Insight Into

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Oxidative stress: a common denominator in every pathogenesis

South African General Practitioner ◽

10.36303/sagp.2020.1.5.0043 ◽

2020 ◽

pp. 202-203

Author(s):

PA De Valdoleiros

Keyword(s):

Oxidative Stress ◽

Signs And Symptoms ◽

Common Denominator ◽

Biological Processes ◽

Medical Doctors ◽

Insight Into

As medical doctors, we are expected to be the champions of health and the slayers of disease. Essential to the success of this battle is an understanding of, and insight into, the chemistry that allows you the opportunity of reading this article. It is not sufficient to name the parts. It is imperative that we understand how the parts work, the processes that lead to malfunctions, and how these malfunctions appear as signs and symptoms to which we eventually give a label. We have concentrated on the results of biological processes gone awry. It is time to deal with the causes.

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Expression of rat fibroblast growth factor receptor 1 as three splicing variants during kidney development

AJP Renal Physiology ◽

10.1152/ajprenal.1993.264.1.f66 ◽

1993 ◽

Vol 264 (1) ◽

pp. F66-F73

Author(s):

E. G. Kim ◽

H. M. Kwon ◽

C. R. Burrow ◽

B. J. Ballermann

Keyword(s):

Kidney Development ◽

Rat Kidney ◽

Cdna Probe ◽

Growth Factor Receptor ◽

Neonatal Rat ◽

Fibroblast Growth ◽

Fgf Receptor ◽

Produce Cell ◽

Splicing Variant ◽

Splicing Variants

Fibroblast growth factors (FGF) are known to participate in the processes of embryogenesis and angiogenesis. This study was undertaken to examine the transcriptional and posttranscriptional regulation of the FGF receptor 1 (FGFR-1) subclass in the embryonic rat kidney. Two full-length FGF receptor cDNAs were cloned using low-stringency screening of a neonatal rat kidney library with a chicken FGFR-1 cDNA probe. Sequencing revealed these cloned cDNAs to be rat homologues of the FGFR-1 subtype, with the two clones representing splicing variants beta and gamma of the FGFR-1. Evidence for renal expression of a third splicing variant (alpha) was obtained by use of the polymerase chain reaction. Splicing variants alpha and beta of FGFR-1 are predicted to produce cell-surface FGF receptors with three and two immunoglobulin-like domains, respectively, whereas the gamma-isoform may represent an intracellular form of the receptor. Although all three splicing variants were expressed in the developing kidney at days 14, 17, and 20 of gestation, at neonatal days 1 and 7 and in mature rats the beta-isoform was present in vastly larger abundance than alpha- and gamma-isoforms at all stages studied. Northern blot analysis revealed enhanced expression of FGFR-1 in the neonatal compared with the mature kidney. It is concluded that FGFR-1 is expressed in the kidney predominantly as the beta-isoform splicing variant and that expression of this receptor is enhanced during kidney development.

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Dynamics Studies of DNA with Non-canonical Structure Using NMR Spectroscopy

International Journal of Molecular Sciences ◽

10.3390/ijms21082673 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2673 ◽

Cited By ~ 1

Author(s):

Kwang-Im Oh ◽

Jinwoo Kim ◽

Chin-Ju Park ◽

Joon-Hwa Lee

Keyword(s):

Nmr Spectroscopy ◽

Biological Function ◽

Conformational Exchange ◽

Biological Processes ◽

Dynamic Characterization ◽

Dynamic Features ◽

Dna Structures ◽

Left Handed ◽

G Quadruplex ◽

Insight Into

The non-canonical structures of nucleic acids are essential for their diverse functions during various biological processes. These non-canonical structures can undergo conformational exchange among multiple structural states. Data on their dynamics can illustrate conformational transitions that play important roles in folding, stability, and biological function. Here, we discuss several examples of the non-canonical structures of DNA focusing on their dynamic characterization by NMR spectroscopy: (1) G-quadruplex structures and their complexes with target proteins; (2) i-motif structures and their complexes with proteins; (3) triplex structures; (4) left-handed Z-DNAs and their complexes with various Z-DNA binding proteins. This review provides insight into how the dynamic features of non-canonical DNA structures contribute to essential biological processes.

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