Phenotypic characterization of SETD3 knockout Drosophila

Tiebe, Marcel; Lutz, Martina S.; Levy, Dan; Teleman, Aurelio A.

doi:10.1371/journal.pone.0201609

Cited by 6 publications

(8 citation statements)

References 51 publications

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“…We next took advantage of the recently published knock-out model of SETD3 in D. melanogaster ( Tiebe et al, 2018 ) to investigate the methylation status of actin in vivo. The absence of SETD3 in the fly prevented the methylation of Drosophila actin on H74 (which corresponds to the H73 in the mammalian protein).…”

Section: Resultsmentioning

confidence: 99%

“…To investigate whether the absence of SETD3 in D. melanogaster also prevented actin methylation, we prepared protein extracts by homogenizing approximately 20 larvae from wildtype or SETD3 KO flies ( Tiebe et al, 2018 ) in 0.5 ml of 25 mM Hepes pH 7.5 with 2.5 µg/ml of leupeptin and antipain. After centrifugation (15 min at 16 000 ×g), the supernatant was recovered, protein concentration was measured ( Bradford, 1976 ) and 10 µg of total proteins were digested with trypsin and used for LC-MS/MS analysis.…”

Section: Methodsmentioning

confidence: 99%

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SETD3 protein is the actin-specific histidine N-methyltransferase

Kwiatkowski

Seliga

Vertommen

et al. 2018

eLife

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140

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Protein histidine methylation is a rare post-translational modification of unknown biochemical importance. In vertebrates, only a few methylhistidine-containing proteins have been reported, including β-actin as an essential example. The evolutionary conserved methylation of β-actin H73 is catalyzed by an as yet unknown histidine N-methyltransferase. We report here that the protein SETD3 is the actin-specific histidine N-methyltransferase. In vitro, recombinant rat and human SETD3 methylated β-actin at H73. Knocking-out SETD3 in both human HAP1 cells and in Drosophila melanogaster resulted in the absence of methylation at β-actin H73 in vivo, whereas β-actin from wildtype cells or flies was > 90% methylated. As a consequence, we show that Setd3-deficient HAP1 cells have less cellular F-actin and an increased glycolytic phenotype. In conclusion, by identifying SETD3 as the actin-specific histidine N-methyltransferase, our work pioneers new research into the possible role of this modification in health and disease and questions the substrate specificity of SET-domain-containing enzymes.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

SETD3 protein is the actin-specific histidine N-methyltransferase

Kwiatkowski

Seliga

Vertommen

et al. 2018

eLife

Self Cite

140

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“…Sztal and Stainer reviewed the genetic compensation response and posed the question of if this mechanism is conserved across species [ 65 ]. Although much of the research discussed herein has employed zebrafish to understand this transcriptional adaptation, several other model organisms, including mouse [ 66 , 67 , 68 ], Drosophila [ 69 , 70 , 71 ], yeast [ 72 , 73 , 74 ], and Arabidopsis [ 75 , 76 , 77 , 78 ], report similar discrepancies which would benefit from in-depth compensation analyses. Additionally, this compensation phenomenon was also evidenced in Caenorhabditis elegans , which also involved nonsense-mediated mRNA decay among other important factors of Argonaute proteins and Dicer [ 79 ].…”

Section: Mechanisms For Genetic Compensationmentioning

confidence: 99%

Genotype to Phenotype: CRISPR Gene Editing Reveals Genetic Compensation as a Mechanism for Phenotypic Disjunction of Morphants and Mutants

Salanga

2021

IJMS

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Forward genetic screens have shown the consequences of deleterious mutations; however, they are best suited for model organisms with fast reproductive rates and large broods. Furthermore, investigators must faithfully identify changes in phenotype, even if subtle, to realize the full benefit of the screen. Reverse genetic approaches also probe genotype to phenotype relationships, except that the genetic targets are predefined. Until recently, reverse genetic approaches relied on non-genomic gene silencing or the relatively inefficient, homology-dependent gene targeting for loss-of-function generation. Fortunately, the flexibility and simplicity of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system has revolutionized reverse genetics, allowing for the precise mutagenesis of virtually any gene in any organism at will. The successful integration of insertions/deletions (INDELs) and nonsense mutations that would, at face value, produce the expected loss-of-function phenotype, have been shown to have little to no effect, even if other methods of gene silencing demonstrate robust loss-of-function consequences. The disjunction between outcomes has raised important questions about our understanding of genotype to phenotype and highlights the capacity for compensation in the central dogma. This review describes recent studies in which genomic compensation appears to be at play, discusses the possible compensation mechanisms, and considers elements important for robust gene loss-of-function studies.

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“…Methylation can modulate protein activity, stability, localization, and/or interactions, resulting in specific downstream signaling and biological outcomes. Lysine methylation is a dynamic and fine-tuned process, and its irregularities often lead to human pathology ( 84 ). HIF1α and HIF2α are the main regulators of cellular responses to hypoxia.…”

Section: Role Of Different Post-translational Modifications In High-a...mentioning

confidence: 99%

The role of post-translational modifications in driving abnormal cardiovascular complications at high altitude

Hou

Wen²,

Long

et al. 2022

Front. Cardiovasc. Med.

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The high-altitude environment is characterized by hypobaric hypoxia, low temperatures, low humidity, and high radiation, which is a natural challenge for lowland residents entering. Previous studies have confirmed the acute and chronic effects of high altitude on the cardiovascular systems of lowlanders. Abnormal cardiovascular complications, including pulmonary edema, cardiac hypertrophy and pulmonary arterial hypertension were commonly explored. Effective evaluation of cardiovascular adaptive response in high altitude can provide a basis for early warning, prevention, diagnosis, and treatment of altitude diseases. At present, post-translational modifications (PTMs) of proteins are a key step to regulate their biological functions and dynamic interactions with other molecules. This process is regulated by countless enzymes called “writer, reader, and eraser,” and the performance is precisely controlled. Mutations and abnormal expression of these enzymes or their substrates have been implicated in the pathogenesis of cardiovascular diseases associated with high altitude. Although PTMs play an important regulatory role in key processes such as oxidative stress, apoptosis, proliferation, and hypoxia response, little attention has been paid to abnormal cardiovascular response at high altitude. Here, we reviewed the roles of PTMs in driving abnormal cardiovascular complications at high altitude.

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Phenotypic characterization of SETD3 knockout Drosophila

Cited by 6 publications

References 51 publications

SETD3 protein is the actin-specific histidine N-methyltransferase

SETD3 protein is the actin-specific histidine N-methyltransferase

Genotype to Phenotype: CRISPR Gene Editing Reveals Genetic Compensation as a Mechanism for Phenotypic Disjunction of Morphants and Mutants

The role of post-translational modifications in driving abnormal cardiovascular complications at high altitude

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