The fail-safe mechanism of post-transcriptional silencing of unsplicedHAC1mRNA.
HAC1encodes a transcription factor that is the central effector of the unfolded protein response (UPR) in budding yeast. When the UPR is inactive,HAC1mRNA is stored as an unspliced isoform in the cytoplasm and no Hac1 protein is detectable. Intron removal is both necessary and sufficient to relieve the post-transcriptional silencing ofHAC1mRNA, yet the precise mechanism by which the intron prevents Hac1 protein accumulation has remained elusive. Here, we show that a combination of inhibited translation initiation and accelerated protein degradation-both dependent on the intron-prevents the accumulation of Hac1 protein when the UPR is inactive. Functionally, both components of this fail-safe silencing mechanism are required to prevent ectopic production of Hac1 protein and concomitant activation of the UPR. Our results provide a mechanistic understanding ofHAC1regulation and reveal a novel strategy for complete post-transcriptional silencing of a cytoplasmic mRNA.
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
Rachael
R
Di Santo
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
Soufiane
S
Aboulhouda
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.|Sandler Faculty Fellows Program, University of California, San Francisco, San Francisco, United States.
David E
0000-0002-9348-1709
DE
Weinberg
D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D007438:Introns; D010442:Peptide Chain Initiation, Translational; D059748:Proteolysis; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D056811:Unfolded Protein Response
D050976:Basic-Leucine Zipper Transcription Factors; C091341:HAC1 protein, S cerevisiae; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins
S. cerevisiae; chromosomes; genes; translation; ubiquitination; unfolded protein response
Philos Trans R Soc Lond B Biol Sci. 1999 Sep 29;354(1389):1533-50
10582239
Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4660-5
10781071
Mol Cell Biol. 2001 Feb;21(3):916-27
11154278
Cell. 2001 Oct 5;107(1):103-14
11595189
Methods Enzymol. 2002;350:87-96
12073338
Biochem J. 2002 Oct 1;367(Pt 1):1-11
12117416
J Biol Chem. 2003 Mar 28;278(13):11441-8
12522142
Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):3889-94
12660367
Nat Rev Mol Cell Biol. 2004 Oct;5(10):827-35
15459663
Nature. 2006 Mar 23;440(7083):561-4
16554824
Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13004-9
16916930
Nature. 2007 Jun 14;447(7146):875-8
17507927
FEBS Lett. 2008 Feb 20;582(4):503-9
18206654
Mol Cell Biol. 2008 Jul;28(13):4227-39
18458056
Science. 2009 Apr 10;324(5924):218-23
19213877
Nat Methods. 2009 May;6(5):343-5
19363495
Cell. 1991 Jan 11;64(1):13-5
1986863
Mol Biol Cell. 2010 Nov 1;21(21):3722-34
20844078
BMC Genomics. 2011 Jan 26;12:68
21269496
Nucleic Acids Res. 2012 Jan;40(Database issue):D700-5
22110037
Nucleic Acids Res. 2012 Apr;40(7):3143-51
22140113
Nucleic Acids Res. 2012 Aug;40(14):6508-19
22544605
Mol Microbiol. 2012 Oct;86(1):225-39
22882838
Cold Spring Harb Protoc. 2012 Oct 01;2012(10):1082-6
23028070
Cold Spring Harb Perspect Biol. 2013 Mar 01;5(3):a013169
23388626
Nat Rev Mol Cell Biol. 2013 Jun;14(6):369-81
23657496
Nature. 2014 Jan 30;505(7485):701-5
24336214
J Biotechnol. 2014 Dec 20;192 Pt B:383-92
24486029
Nucleic Acids Res. 2014;42(17):e134
25056308
J Biol Chem. 2015 Sep 4;290(36):21821-32
26175153
Cell. 2015 Oct 8;163(2):292-300
26451481
Cell Rep. 2016 Feb 23;14(7):1787-1799
26876183
Nucleic Acids Res. 2016 Jul 8;44(W1):W3-W10
27137889
Bio Protoc. 2017 Oct 5;7(19):null
29170751
EMBO J. 1983;2(12):2161-8
6321150
Cell. 1993 Jul 30;74(2):357-69
8393731
Cell. 1996 Nov 1;87(3):391-404
8898193
Cell. 1996 Nov 1;87(3):405-13
8898194
Nucleic Acids Res. 1996 Nov 1;24(21):4222-6
8932376
Genes Cells. 1996 Sep;1(9):803-17
9077435
Cell. 1997 Sep 19;90(6):1031-9
9323131
Mol Biol Cell. 1997 Oct;8(10):1845-62
9348528
Curr Biol. 1997 Nov 1;7(11):850-9
9382810
J Biotechnol. 1998 Jun 11;62(1):29-45
9684340
Annu Rev Cell Dev Biol. 1998;14:459-85
9891790