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MBB331_Week10-Fall-2011

2 phase 1 pap adds 10 phase polya binding protein

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Unformatted text preview: CPSF BINDING TO THE AAUAAA MOTIF CPSF BINDING TO THE AAUAAA MOTIF • Labeled Labeled oligoribooligoribonucleotide nucleotide mixed with poly(A)polypoly(A)polymerase or merase CPSF CPSF • CPSF binds CPSF w.t. but not mutant oligos oligos • Proteins Xlinked to linked oligos (by u.v.) show major CPSF complexes complexes Weaver 2002 Molecular Biology Ed. 2 PAP = poly(A)polymerase CPSF = specificity component PAP 60 3’ PROCESSING COMPLEX 3’ PROCESSING COMPLEX • • • CPSF complexes with CPSF endonuclease (CFI & CFII) & poly(A) polymerase (PAP) poly(A) endonuclease cleaves endonuclease transcript downstream of AAUAAA, and PAP synthesizes poly(A) tail poly(A) PAP has nonspecific catalytic PAP activity which when combined with other components becomes specific for RNA containing AAUAAA containing Weaver 2002 Molecular Biology Ed. 2 Phase 1: PAP adds 10 Phase polyAs polyAs Phase 2: PABP Phase promotes polyA tail extension to ~200 A’s extension 61 3’ PROCESSING COMPLEX 3’ PROCESSING COMPLEX Weaver 2002 Molecular Biology Ed. 2 Phase 1: PAP adds 10 Phase • • Poly(A) binding protein (PABP) Poly(A) associates with tail and aids PAP in elongating tail to >200 A’s A’s PABP remain...
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