Dataset Information

Phosphoproteomic screening identifies physiological substrates of dTSSK kinase

ABSTRACT: TO identified its potential phosphorylated substrates by investigating differential phosphorylated sites in the testes of wild-type (w1118) and dTSSK-/- male flies using quantitative phosphoproteomic analysis

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Testis

SUBMITTER: Xuedi Zhang

LAB HEAD: Guanjun Gao

PROVIDER: PXD038767 | Pride | 2023-07-20

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Dj15ZXD_bjhb1_20201120001855.raw	Raw
	Dj15ZXD_bjhb1_pho__2_.pep.xml	Pepxml
	Dj15ZXD_bjhb1_pho__2_.prot.xml	Xml

Items per page:

1 - 3 of 3

Publications

Broad phosphorylation mediated by testis-specific serine/threonine kinases contributes to spermiogenesis and male fertility.

Zhang Xuedi X Peng Ju J Wu Menghua M Sun Angyang A Wu Xiangyu X Zheng Jie J Shi Wangfei W Gao Guanjun G

Nature communications 20230506 1

Genetic studies elucidate a link between testis-specific serine/threonine kinases (TSSKs) and male infertility in mammals, but the underlying mechanisms are unclear. Here, we identify a TSSK homolog in Drosophila, CG14305 (termed dTSSK), whose mutation impairs the histone-to-protamine transition during spermiogenesis and causes multiple phenotypic defects in nuclear shaping, DNA condensation, and flagellar organization in spermatids. Genetic analysis demonstrates that kinase catalytic activity o ...[more]

PMID: 37149634

Similar Datasets

Project description:1α,25(OH)2VD3 is the most active form of VD3 in animals and it plays an important role in regulating mineral metabolism but also reproduction and immunity. Testes are the main reproductive organs of male mammals. In order to study the effect of 1α,25(OH)2VD3 on early testicular development, 140 weaned 21-day old piglets were selected. The piglets were randomly divided into four groups and were fed a commercial diet supplemented with 0, 1, 2 and 4 μg.kg-1 of 1α,25(OH)2VD3, provided as 1α,25(OH)2VD3-glycosides, 60 days after the start of the experiment, at piglet age 82 days, testes were harvested. The morphology and histology of early testicular development were assessed. In addition, the proteomic TMT/iTRAQ labeling technique was used to analyze the protein profile of the testes in each group. Western blotting was used to verify the target of differentially abundant proteins (DAPs). The results showed that of the identified 132715 peptides, 122755 were specific peptides. 7852 proteins, of which 6573 proteins contain quantitative information. Screening for DAPs was focused on proteins closely related to the regulation of the testicular development such as steroid hormone synthesis, steroid biosynthesis, peroxisome and fatty acid metabolism pathways. These results also indicate that 1α,25(OH)2VD3 is involved in the regulation of early testicular development in piglets. At the same time, these findings provide valuable information for the proteins involved in the regulation of testicular development, and help to better understand the mechanisms of 1α,25(OH)2VD3 in regulating the development of piglets’ testes. Significance: The early development of the testes of breeding boars is closely related to their lifetime fertility. Our research identifies potential signaling pathways and differentially abundant proteins associated with testicular development. These findings indicate that plants containing 1α,25(OH)2VD3-glycosides used as a nutritional ingredient may help early testicular development in breeding boars and reveal the regulatory mechanism of 1α,25(OH)2VD3 on testicular development.

Project description:Environmental reproductive health focuses on the effect of exposure to contaminants considered as endocrine disruptors. Developmental testis is considered as target of these compounds affecting testicular functions in adults and suspected implications in tumor etiology. Comparative analysis of gene expression in mouse testis exposed to five disruptors, three different dosages and three accumulative developmental stages shown defined signature profiles of gene deregulation for MEHP (monoethyl phthalate) and zearalenone (a phytoestrogen) and different to 17β-estradiol exposure. The effects are even detected in postpuberal male offspring from premating exposed mothers. Oxidative stress response, protein ubiquitination and oxidative phosphorylation are the most representative pathways affected. Animal Exposure CD-1 mice were in vivo exposed to several doses of EDs following a defined protocol: mothers were exposed two weeks before mating (exposure A); the same exposure and dose were maintained during pregnancy (exposure B) and four weeks after birth (exposure C). In all exposures, male offspring were sacrificed to obtain the testes. The compounds were administrated in drinking water to a final concentration of: E2(mg/l) BPA(mg/l) Zea(mg/l) MEHP(mg/l) Lindane(mg/l) Dose (1) 0.02 0.5 4 27.85 50 Dose (2) 0.04 50 12 139.17 100 Dose (3) 0.16 200 20 278.34 200 Ethanol was used as vehicle for E2, BPA and Zea and DMSO was for MEHP and Lindane. Control testes were obtained from animals exposed to solvents of EDs: Ethanol and DMSO. Animal care and sacrifice were made following regulations from the CSIC Bioethics Committee. RNA preparation Total RNA from testes of exposed animals and controls was extracted using TRIzol reagent (Invitrogen), according to the manufacturer’s standard protocol. RNA samples were spectrophotometrically quantified using NanoDrop (NanoDrop Technologies). To minimize the effect of inter-individual expression variability we made pools of RNA from at least 3 individual RNA samples from each experimental condition (same quantity of total RNA per sample). Pools of RNA were checked for quality performing Experion analysis (Bio-Rad Laboratories). Total RNA from testes of unexposed four weeks old animals was also extracted with the purpose of making pools to assess the variability at the expression level of the mouse strain used (Ctr). Microarray Processing Starting with total RNA from pools described above, full length cDNAs were synthesized from oligo(dT) primers bearing a T7 promoter. These cDNAs were used as template for in vitro transcription by T7-RNA-Polymerase to amplify the mRNAs; following the Amino Allyl MessageAmp™ aRNA Kit (Ambion, Inc.) protocol. The amplified mRNAs (aRNA) were labeled either with Cy3 or Cy5 dyes, using Amersham CyDye Post-Labeling Reactive Dyes (Amersham Biosciences). The incorporated dye was spectrophotometrically quantified using NanoDrop (NanoDrop Technologies). The same dye signal quantities of labeled experimental and control aRNAs were mixed and hybridized to Mouse Oligoset v3 (OPERON) arrays from Genomics facility-University of Cincinnati (USA). The slides contained 31,769 spotted probes (70 mer oligonucleotides) corresponding to 24,878 expressed or predicted mouse genes. Hybridization and washes of microarrays were performed according to Genomics facility-University of Cincinnati instructions. Dye-Swap replicates were performed: one slide hybridized with experimental and control targets labeled with Cy3 and Cy5, respectively; and the second slide with the same type of targets but inversely labeled. Target refers to labeled aRNA while probe refers to the oligonucleotides spotted on the microarray platform. Imaging and data generation were carried out using a GenePix 4000B (Axon Instruments) and associated software from Axon Instruments, Inc. The microarray slides were scanned with dual lasers with wavelength frequencies to excite Cy3 and Cy5. Images were captured in TIFF files. Information extraction for a given spot was based on the median value for the signal pixels minus the median value for the background pixels to produce a gene set data file for all the DNA spots. The Cy3 and Cy5 fluorescence signal intensities were normalized.