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MAP kinase Pathways in Yeast

MAP kinase Pathways in Yeast. How are signals specified between pathways that share Common components? MAP = mitogen activated protein kinase. FGP ? Cdc42/Ste20 Ste11 Ste7 Kss1 Ste12/ Tec1. PRP Ste2/3/4/18 Cdc42/Ste20 Ste11 Ste7 Fus3 Ste12. Ste5. Two MAP kinase pathways.

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MAP kinase Pathways in Yeast

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  1. MAP kinase Pathways in Yeast How are signals specified between pathways that share Common components? MAP = mitogen activated protein kinase

  2. FGP ? Cdc42/Ste20 Ste11 Ste7 Kss1 Ste12/Tec1 PRP Ste2/3/4/18 Cdc42/Ste20 Ste11 Ste7 Fus3 Ste12 Ste5 Two MAP kinase pathways

  3. Two MAP kinase pathways

  4. UBIQUITIN • 76 Amino Acid polypeptide • Highly conserved in evolution: 3 Amino acid differences between yeast and human homologues

  5. Degradation of a Protein Via the Ubiquitin-Proteasome Involves Two Successive Steps • 1. Covalent attachment of multiple ubiquitin molecules to a protein substrate. • 2. Degradation of the tagged protein by the 26s proteasome. (ubiquitin is recycled)

  6. UBIQUITINATION In general, multiple ubiquitin units are arranged in polyubiquitin chains linked via Lys48 of ubiquitin, targeting the protein for degradation

  7. Ubiquitin Conjugation: A 3 Step Mechanism

  8. Ub Ub The Ubiquitin Pathway Ub E1 E2 E3 Target Ub Ub Ub Ub 26s proteosome degradation

  9. Classes of E3 ligases • N-end Rule E3 • Skp/Cullin/F-box (SCF) • Anaphase Promoting Complex • HECT family • RING finger domain

  10. Phosphorylation of I leads to ubiquitination and degradation of the inhibitor and activation of the transcription factor.

  11. ATP consuming process

  12. THE PROTEASOME

  13. Small Ubiquitin Related Modifier (SUMO) • SUMO does not have the Lys-48 found in ubiquitin • SUMO does not make multi-chain forms • SUMO-1,2,3 are the mammalian forms • SUMO-1, 101 amino acids, C-terminal Gly, 18% identical to ubiquitin.

  14. SUMO

  15. MASS-SPEC AnalysisAllows determination of peptide sequences and modified peptides (by P~)Mass spectrometers separate gas-phase ions according to their mass to charge ratiosThe analyzer uses a combination of magnetic and electric fields to move ions from the regions that they are produced to the detector.

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