Dataset Information

Phosphoproteomics identifies targets of Mos-MAPK regulating translation and spindle organization in oocyte meiosis

ABSTRACT: The Mos kinase is a constitutive activator of the ERK/MAPK pathway exclusively expressed during oocyte meiosis, mediating key meiotic functions across animal species. While a few of its downstream effectors have been studied in some detail, molecular targets under the control of Mos-MAPK have not yet been identified systematically. Here, we combined live-cell microscopy of starfish oocytes to characterize the cellular phenotypes caused by Mos-MAPK inhibition with phosphoproteomic analysis of synchronous oocyte populations at critical transitions. This revealed a large set of proteins involved in regulation of translation through the CPE-element binding protein CPEB. Our data indicate cyclin B to be a main target of this regulation driving the second meiotic division. A second large group of phospho-proteins identified are regulators of the actin and microtubule cytoskeleton, with a prominent subset of regulators of centrosomal microtubule nucleation. Indeed, we show that Mos-MAPK inhibition increases the size of microtubule asters and promotes separation of spindle poles in anaphase, turning meiotic spindles mitotic-like. We thus identified core molecular modules downstream of Mos-MAPK controlling meiotic functions essential for haploidization and for the highly asymmetric polar body divisions. These modules are widely conserved, thus our findings will likely have general relevance for reproductive processes across species, including humans, and for understanding disease mechanisms when Mos is expressed erroneously, acting as an oncogene.

INSTRUMENT(S):

ORGANISM(S): Echinoderma Sp.

TISSUE(S): Oocyte

SUBMITTER: Olexandr Dybkov

LAB HEAD: Henning Urlaub

PROVIDER: PXD043650 | Pride | 2025-09-03

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	LWelp_TD_IAvilov_011121_151121_Starfish_oocytes_TMT_Fullprot_bRP1.raw	Raw
	LWelp_TD_IAvilov_011121_151121_Starfish_oocytes_TMT_Fullprot_bRP2.raw	Raw
	LWelp_TD_IAvilov_011121_151121_Starfish_oocytes_TMT_Fullprot_bRP3.raw	Raw
	LWelp_TD_IAvilov_011121_151121_Starfish_oocytes_TMT_Fullprot_bRP4.raw	Raw
	LWelp_TD_IAvilov_011121_151121_Starfish_oocytes_TMT_Fullprot_bRP5.raw	Raw

Items per page:

1 - 5 of 49

Similar Datasets

Project description:The identification of genes and deduced pathways from the mature human oocyte can help us better understand oogenesis, folliculogenesis, fertilization, and embryonic development. Human metaphase II oocytes were used within minutes after removal from the ovary, and its transcriptome was compared with a reference sample consisting of a mixture of total RNA from 10 different normal human tissues not including the ovary. RNA amplification was performed by using a unique protocol. Affymetrix Human Genome U133 Plus 2.0 GeneChip arrays were used for hybridizations. Compared with reference samples, there were 5,331 transcripts significantly up-regulated and 7,074 transcripts significantly down-regulated in the oocyte. Of the oocyte up-regulated probe sets, 1,430 have unknown function. A core group of 66 transcripts was identified by intersecting significantly up-regulated genes of the human oocyte with those from the mouse oocyte and from human and mouse embryonic stem cells. GeneChip array results were validated using RT-PCR in a selected set of oocyte-specific genes. Within the up-regulated probe sets, the top overrepresented categories were related to RNA and protein metabolism, followed by DNA metabolism and chromatin modification. This report provides a comprehensive expression baseline of genes expressed in in vivo matured human oocytes. Further understanding of the biological role of these genes may expand our knowledge on meiotic cell cycle, fertilization, chromatin remodeling, lineage commitment, pluripotency, tissue regeneration, and morphogenesis. Experiment Overall Design: We generated a database of the human oocyte transcriptome by comparing the transcripts in the oocyte with the reference samples, which contain mRNA from several somatic tissues. Experiment Overall Design: Human oocytes were obtained from three patients undergoing an assisted reproductive treatment (ART) at the Unit of Reproductive Medicine of Clinica Las Condes, Santiago, Chile. It is important to emphasize that the routine in vitro fertilization protocol at Clinica Las Condes calls for fertilizing only those oocytes that will be transferred into the uterus of the patient. Therefore, there is always a surplus of oocytes. We then had the opportunity to use specific criteria to select donors as follows: (i) <35 years old, (ii) reproductively healthy with regular ovulatory cycles, (iii) male factor as the only cause of infertility, and (iv) considerable number of developing follicles that assured spared oocytes. The experimental protocol was reviewed and approved by a local independent Ethics Review Board. All donors signed informed consent. At the time this manuscript was submitted, all three donors had already conceived; two of them got pregnant during the ART cycle in which our samples were collected, and the third one got pregnant after a spontaneous cycle with artificial insemination using donated sperm. Ovarian stimulation, oocyte retrieval, and cell lysis were performed as described in Supporting Materials and Methods, which is published as supporting information on the PNAS web site. Experiment Overall Design: Three groups of 10 oocytes each were used. Reference RNA (100 Î¼g) was prepared by mixing 10 Î¼g of total RNA from each of 10 different normal human tissues, including skeletal muscle, kidney, lung, colon, liver, spleen, breast, brain, heart, and stomach (Ambion). Experiment Overall Design: Transcriptional profile of each sample was probed by using Affymetrix Human Genome U133 Plus 2.0 GeneChips.

Dataset Information

Phosphoproteomics identifies targets of Mos-MAPK regulating translation and spindle organization in oocyte meiosis

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets