The mRNA encoding the yeast ARE-binding protein Cth2 is generated by a novel 3' processing pathway.

Nucleic Acids Research
Delphine CiaisDavid Tollervey

Abstract

Microarray analyses of mRNAs over-expressed in strains lacking the nuclear exosome component Rrp6 identified the transcript encoding the ARE-binding protein Cth2, which functions in cytoplasmic mRNA stability. Subsequent northern analyses revealed that exosome mutants accumulate a 3'-extended transcript at the expense of the mature CTH2 mRNA. The 3' ends of the CTH2 mRNA were mapped to a [GU(3-5)](5) repeat, unlike any previously characterized polyadenylation site. CTH2 mRNA accumulation was not inhibited by mutations in 3'-cleavage and polyadenylation factors, Rna14, Rna15 and Pap1, which block accumulation of other mRNAs. The 3'-extended CTH2 pre-mRNA strongly accumulated in strains with mutations in the TRAMP4 polyadenylation complex or the Nrd1/Nab3/Sen1 complex, and contains multiple Nrd1 and Nab3 binding sites. CTH2 carries a consensus ARE element and levels of the pre-mRNA and mRNA were elevated by mutation of the ARE or inactivation of the nuclear 5'-exonuclease Rat1. We propose that CTH2 mRNA is processed from a 3'-extended primary transcript by the exosome, TRAMP and Nrd1/Nab3/Sen1 complexes. This unusual pathway may allow time for nuclear, ARE-mediated regulation of CTH2 levels involving Rat1.

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Citations

Aug 6, 2013·Trends in Biochemical Sciences·Claudia Schneider, David Tollervey
Aug 19, 2009·Molecular and Cellular Biology·Yung-Shu KuanLillie L Searles
Dec 17, 2009·RNA·Jeremy E Wilusz, David L Spector
Aug 2, 2013·International Journal of Molecular Sciences·María Teresa Martínez-PastorSergi Puig
Dec 14, 2011·Trends in Genetics : TIG·Oliver Mühlemann, Torben Heick Jensen
Sep 8, 2015·The Journal of Nutritional Biochemistry·Amanda J Bird
Oct 17, 2008·The EMBO Journal·Manoël ProuteauBertrand Séraphin
Apr 5, 2011·The EMBO Journal·Wiebke WlotzkaDavid Tollervey
Apr 17, 2012·The EMBO Journal·Rajani Kanth GudipatiAlain Jacquier
Feb 19, 2013·Wiley Interdisciplinary Reviews. RNA·Karyn Schmidt, J Scott Butler
May 27, 2014·Nucleic Acids Research·Veronika BacikovaRichard Stefl
Feb 5, 2013·Biochemical and Biophysical Research Communications·Koichi OgamiShin-ichi Hoshino
Oct 23, 2012·Biochimica Et Biophysica Acta·Hannah E Mischo, Nick J Proudfoot
Apr 10, 2018·Non-coding RNA·Koichi OgamiJames L Manley
May 6, 2019·Microbiology·Frédéric Devaux, Antonin Thiébaut
May 27, 2014·RNA·Borislava TsanovaAmbro van Hoof

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Methods Mentioned

BETA
GTPase
PCR

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