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Function of the Ski4p (Csl4p) and Ski7p Proteins in 3’-to-5’ Degradation of mRNA

Function of the Ski4p (Csl4p) and Ski7p Proteins in 3’-to-5’ Degradation of mRNA. Ambro van Hoof, Robin R. Staples, Richard E. Baker, and Roy Parker. Molcecular and Cellular Biology , p8230-8243 (2000). Introduction. Yeast virus. Annu. Rev. Genet . 30:109-39 (1996).

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Function of the Ski4p (Csl4p) and Ski7p Proteins in 3’-to-5’ Degradation of mRNA

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  1. Function of the Ski4p (Csl4p) and Ski7p Proteins in 3’-to-5’ Degradation of mRNA Ambro van Hoof, Robin R. Staples, Richard E. Baker, and Roy Parker Molcecular and Cellular Biology, p8230-8243 (2000)

  2. Introduction Yeast virus Annu. Rev. Genet. 30:109-39 (1996)

  3. Annu. Rev. Microbiol. 46:347-75 (1992) Crystal structure 3.4Å Cryo-EM 16Å J. Mol. Recognit. 18:158-168 (2005)

  4. The viral replication cycles of L-A and its satellites J. Cell. Biol. 138:975-985 (1997 ) Annu. Rev. Genet. 30:109-39 (1996)

  5. SKI antiviral system J Bacteriol. 136(3): 1002–1007 (1978) Trends. Microbiol. 1:294-298 (1993)

  6. p 7mGppp 7mGppp A60-80 A60-80 A0-10 mRNA mRNA mRNA mRNA degradation in yeast Deadenylation Decapping (Dcp1p, Dcp2p) 3’-5’ degradation 5’-3’ degradation (Xrn1p) 7mGppp mRNA A60-80 mRNA The exosome: a conserved eukaryotic RNA processing complex containing multiple 3’-5’ exoribonuclease activities. Cell, 91, 457-466 (1997) Rrp4p,Rrp41/Ski6p, Rrp42p, Rrp43p, Rrp44p

  7. The 3’ to 5’ degradation of yeast mRNAs is a general mechanism for mRNA turnover that requires the SKI2 DEVH box protein and 3’ to 5’ exonucleases of the exosome complex EMBO J. 1497-1506 (1998) The SKI2, SKI3, SKI6/RRP41, SKI8 and RRP4 gene products are required for 3’ to 5’ decay of mRNA The yeast antiviral proteins Ski2p, Ski3p, and Ski8p exist as a complex in vivo RNA 6:449-457 (2000)

  8. The ski4-1 and ski7Δ mutations affect the metabolism of a degradation intermediate of MFA2pG mRNA

  9. The ski4-1 and ski7Δ mutations stabilize a degradation intermediate of MFA2pG mRNA

  10. Strains containing either ski4-1 or ski7Δ in combination with dcp1-2 or dcp2-7 are not able to grow under conditions restrictive for the decapping defect SKi4p and Ski7p proteins are required for 3’-5’ mRNA degradation as are the components of the exosome and the Ski2p,SKi3p, and Ski8p protein.

  11. The ski4-1 mutation is complemented by a wild-type (WT) CSl4 gene

  12. A mutation in the conserved S1 RNA binding domain of Csl4p is responsible for the ski4-1 phenotype Two mutations of Ski4p : R65K, G253E The SKI4 gene is identical to the CSl4 gene

  13. The ski4-1 mutation does not affect all Csl4p functions ski6

  14. The csl4-1 mutation affects rRNA processing but not mRNA degradation rRNA processing mRNA degradation The ski4-1 allele genetically separates the functions of the exosome in rRNA processing and mRNA decay

  15. The ski7Δ and hgs1Δ mutations do not affect 5.8S rRNA or U4 snRNA processing or 5’ ETS degradation The SKI7 gene is specifically required for mRNA degradation by the exosome

  16. Summary 1. The Ski4p and Ski7p proteins are required for mRNA degradation. 2. The SKI4 gene is identical to the CSL4 gene 3. The ski4-1 allele genetically separates the functions of the exosome in rRNA processing and mRNA decay 4.The SKI7 gene is specifically required for mRNA degradation by the exosome. SKI 1: Xrn1p enzyme (5’ to 3’ exoribonuclease) SKI 2: 1287 a.a. putative RNA helicase SKI 3: 1432 a.a. tetratricopeptide repeat (TRP) protein SKI 4: 195 a.a. S1 domain (Csl4p) SKI 6: 264 a.a. Rrp41p (3’-5’ exoribonuclease) SKI 7: 747 a.a. GTPase domain ( translation elongation factor EF1A/ translation termination factor eRF3) SKI 8: 397 a.a. five WD-40 (beta-transducin) repeats protein

  17. Componenets of the exosome complex

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