Fate of mRNA after RNA processing events in nucleus Hafidh Adv Exp Med Biol. 722:118, 2011.

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Fate of mRNA after RNA processing events in nucleus Hafidh Adv Exp Med Biol. 722:118, 2011

“Proofreading and spellchecking: A two-tier strategy for pre- mRNA splicing quality control” Egecioglu & Chanfreau RNA 17: 383, 2011 DEAD-box helicases (ATPase Prp proteins) unwind incorrect base-pairing What happens if splicing pathway is defective?

Egecioglu & Chanfreau RNA 17: 383, 2011 RNA degradation pathways for RNAs that escaped splicing or have splicing mistakes “Quality control” removal of defective transcripts

Nonsense-mediated mRNA decay (NMD) - surveillance system to remove defective RNAs - mRNAs with premature stop codons are rapidly degraded Gonzalez Gene 274: 15, 2001 Observations: - in “pioneer round” of translation, if EJC (exon joining complex) not removed by ribosomal machinery, it acts as tag for RNA degradation pathway Wilusz Genes & Dev 15:2781, 2001

Steady state levels of RNase-L mRNA quantified by qPCR and normalized relative to constitutively expressed rpl13a mRNA 3’UTR deletion constructs Example of AREs (AU-rich cis-elements) in 3’ UTR of RNase-L mRNA Deletion constructs were transfected into 293T cells and RNase-L expression was monitored by Western blots Li et al. J Biol Chem 282:7950, 2007 Interpretation of data? What experiments might be interesting to do next? Cis-determinants of mRNA stability ZC5+

Kloc Cell 108: 533, 2002 Drosophila egg: nanos mRNA Xenopus oocyte: fatVg mRNA Chicken fibroblast:  -actin mRNA (red) in leading lamellae Mechanisms that target certain mRNAs to appropriate subcellular site? Heredia Curr Opin Cell Biol 16:80, 2004 Localization of mRNAs within eukaryotic cells

Becalska & Gavis Develop 136:2493, 2009 “Lighting up mRNA localization in Drosophila oogenesis”

Advantages of mRNA subcellular localization (and localized translation)? Martin & Ephrussi Cell 136:719, 2009

- mRNAs (eg nanos, bicoid, oscar, K10…) important for developmental processes are synthesized in nurse cells and move to oocyte along microtubules mRNA localization in Drosophila oocyte / embryo Saxton Cell 107:707, 2001 Kloc & Etkin J.Cell Sci. 118:269, 2005 (Fig.4)

Serano Develop. 121: 3809, nt TLS (transport/localization sequence) with stem-loop structure in 3’UTR Model for Drosophila K10 mRNA movement? K  SD

Serano Develop. 121: 3809, 1995 Mutational analysis of stem-loop in 3’ UTR of K10 mRNA

Huang Curr Opin Cell Biol 16:308, 2004 Mechanisms for localized translation of mRNAs