Dataset Information

Yeast meiotic proteome in wildtype versus kar4 mutant

ABSTRACT: The Kar4 protein has been identified as being a component of the yeast N6A mRNA methyltransferase, homologous to human METTL14. Mutations in KAR4 cause blocks in meiosis due to 1) reduced expression of Ime1p-induced transcripts, and 2) reduced levels of a subset of meiotic proteins, independent of their transcript levels. The reduced levels of the proteins are partially suppressed by increased levels of a translational regulator, Rim4. This proteomic study was aimed at identifying the spectrum of meiotic proteins affected by the absence of Kar4. The defect in Ime1p-depedent transcription was suppressed by over-expression of Ime1p. Proteins present in wildtype and kar4 deletion strains at two different times in meiosis were fractionated using reverse-phase chromatography and identified by MS/MS. Some 4068 proteins expressed during meiosis were identified. At 8 hours, 432 proteins were present at less than 50% the levels in kar4Δ/Δ compared to wild type. GO term analysis of the low proteins at 8 hours returned “meiotic cell cycle” as the sixth term with a P value of 7.74x10^-5. At 12 hours, 318 proteins were present at less than 50% the levels in kar4Δ/Δ relative to wild type. GO term analysis returned “meiotic cell cycle” as the second term with a P value less than 0.001. Proteins that were low at both 8 and 12 hours in kar4∆/∆ comprise proteins required for meiotic recombination and sporulation including Sps2p, Gas4p, Gmc2p, Mei5p, and Sae3p. Other meiotic proteins were expressed at or above wild type levels at 8 hours, but went down to lower than wild type levels at 12 hours, including Ecm11p, Hed1p, Spo11p, and Rec8p. In each case the levels of the proteins were reduced to much greater extents that can be explained by changes in transcript levels. Taken together these data suggest that Kar4 plays a role in the regulation of transcription and translation during yeast meiosis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Meiosis Ii

SUBMITTER: Mark Rose

LAB HEAD: Mark D. Rose

PROVIDER: PXD043798 | Pride | 2023-08-22

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	01212016_539_KAR4_12H-2_QE_RPHP1.raw	Raw
	01212016_539_KAR4_12H-2_QE_RPHP2.raw	Raw
	01212016_539_KAR4_12H-2_QE_RPHP3.raw	Raw
	01212016_539_KAR4_12H-2_QE_RPHP4.raw	Raw
	01212016_539_KAR4_12H-2_QE_RPHP5.raw	Raw

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Publications

Kar4 is Required for the Normal Pattern of Meiotic Gene Expression.

Park Zachory M ZM Remillard Matthew M Rose Mark D MD

bioRxiv : the preprint server for biology 20230129

Kar4p, the yeast homolog of the mammalian methyltransferase subunit METTL14, is required for the initiation of meiosis and has at least two distinct functions in regulating the meiotic program. Cells lacking Kar4p can be driven to sporulate by co-overexpressing the master meiotic transcription factor, <i>IME1</i> , and the translational regulator, <i>RIM4</i> , suggesting that Kar4p functions at both the transcriptional and translational level to regulate meiosis. Using microarray analysis and R ...[more]

PMID: 36747654

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Project description:Ray2013 - Meiotic initiation in S. cerevisiae A mathematical representation of early meiotic events, particularly feedback mechanisms at the system level and phosphorylation of signalling molecules for regulating protein activities, is described here This model is described in the article: Dynamic modeling of yeast meiotic initiation. Ray D, Su Y, Ye P. BMC Syst Biol. 2013 May 1;7:37 Abstract: BACKGROUND: Meiosis is the sexual reproduction process common to eukaryotes. The diploid yeast Saccharomyces cerevisiae undergoes meiosis in sporulation medium to form four haploid spores. Initiation of the process is tightly controlled by intricate networks of positive and negative feedback loops. Intriguingly, expression of early meiotic proteins occurs within a narrow time window. Further, sporulation efficiency is strikingly different for yeast strains with distinct mutations or genetic backgrounds. To investigate signal transduction pathways that regulate transient protein expression and sporulation efficiency, we develop a mathematical model using ordinary differential equations. The model describes early meiotic events, particularly feedback mechanisms at the system level and phosphorylation of signaling molecules for regulating protein activities. RESULTS: The mathematical model is capable of simulating the orderly and transient dynamics of meiotic proteins including Ime1, the master regulator of meiotic initiation, and Ime2, a kinase encoded by an early gene. The model is validated by quantitative sporulation phenotypes of single-gene knockouts. Thus, we can use the model to make novel predictions on the cooperation between proteins in the signaling pathway. Virtual perturbations on feedback loops suggest that both positive and negative feedback loops are required to terminate expression of early meiotic proteins. Bifurcation analyses on feedback loops indicate that multiple feedback loops are coordinated to modulate sporulation efficiency. In particular, positive auto-regulation of Ime2 produces a bistable system with a normal meiotic state and a more efficient meiotic state. CONCLUSIONS: By systematically scanning through feedback loops in the mathematical model, we demonstrate that, in yeast, the decisions to terminate protein expression and to sporulate at different efficiencies stem from feedback signals toward the master regulator Ime1 and the early meiotic protein Ime2. We argue that the architecture of meiotic initiation pathway generates a robust mechanism that assures a rapid and complete transition into meiosis. This type of systems-level regulation is a commonly used mechanism controlling developmental programs in yeast and other organisms. Our mathematical model uncovers key regulations that can be manipulated to enhance sporulation efficiency, an important first step in the development of new strategies for producing gametes with high quality and quantity. This model is hosted on BioModels Database and identified by: BIOMD0000000626 . To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models . To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.