Comment[ArrayExpressAccession]	E-GEOD-52929
MAGE-TAB Version	1.1														
Public Release Date	2013-12-04
Investigation Title	Sel1L is Indispensable for Mammalian ERAD, ER Homeostasis and Survival														
Comment[Submitted Name]	Sel1L is Indispensable for Mammalian ERAD, ER Homeostasis and Survival
Experiment Description	Sel1L is an adaptor protein for the E3 ligase Hrd1 involved in endoplasmic reticulum-associated degradation (ERAD). Its physiological importance in mammalian ERAD, however, remains to be established. Here, using the inducible Sel1L knockout mouse and cell models, we provide the first in vivo evidence that Sel1L is indispensable for Hrd1 stability, ER homeostasis and survival. Acute loss of Sel1L leads to premature death in adult mice within 3 weeks with profound pancreatic atrophy. Contrary to current belief, our data show that mammalian Sel1L is required for Hrd1 stability and ERAD function both in vitro and in vivo. Sel1L deficiency disturbs ER homeostasis, activates ER stress, attenuates translation and promotes cell death. Serendipitously, using biochemical approach coupled with mass spectrometry, we found that Sel1L deficiency causes the aggregation of both small and large ribosomal subunits. Thus, Sel1L is an indispensable component of mammalian ERAD and ER homeostasis, which is essential for protein translation, pancreatic function, cellular and organismal survival. Pancreas tissue of wild type and inducible Sel1L knockout mice were subjected to gene expression analysis.														
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl													
Person Last Name	Hooiveld	Sun	Shi	Han	Francisco	Ji	Mendonça	Liu	Locasale	Simpson	Duhamel	Kersten	Yates III	Long	Qi
Person First Name	Guido	Shengyi	Guojun	Xuemei	Adam	Yewei	Nuno	Xiaojing	Jason	Kenneth	Gerald	Sander	John	Qiaoming	Ling
Person Mid Initials					B				W	W			R		
Person Email	guido.hooiveld@wur.nl														
Person Affiliation	Wageningen University														
Person Address	Div. Human Nutrition, Wageningen University, Bomenweg 2, Wageningen, Netherlands														
Person Roles	submitter														
Protocol Name	P-GSE52929-1	P-GSE52929-5	P-GSE52929-6	P-GSE52929-2	P-GSE52929-3	P-GSE52929-4	P-GSE52929-7								
Protocol Description	Expression estimates were calculated applying the RMA algorithm in the Bioconductor library 'Oligo' (v1.24.2). ID_REF =  VALUE = RMA signal (as log2)	One hundred nanogram of RNA was used for whole transcript cDNA synthesis with the Ambion WT expression kit [catalog number 4411974] (Applied Biosystems/Life Technologies, Nieuwekerk a/d IJssel, The Netherlands).	Hybridization and washing of the Affymetrix GeneChip Mouse Gene 1.1 ST peg arrays were performed on a GeneTitan Instrument (Affymetrix, Santa Clara, CA) according to the manufacturer's recommendations.	Tamoxifen (Sigma T5648) was dissolved in sunflower oil at 5 mg/ml. Age-matched littermates at the age of 8-20 weeks were injected intraperitoneally with tamoxifen at the volume of 5 μl/g body weight daily for three consecutive days.	The Sel1ltm1e(KOMP)Wtsi (Sel1Lflox/+) ES cells on the C57BL/6N background were purchased from the KOMP Repository Project (ID CSD44577, UC Davis, http://www.knockoutmouse.org/martsearch/project/44577). Exon 6 of Sel1L gene was flanked with two loxP sites (floxed). The lacZ-neo cassette was deleted by crossing the animals onto the βActin-FLPe deleter mice on the C57BL/6J background (JAX 003800). The resulting Sel1Lflox/+ animals were intercrossed to generate Sel1Lflox/flox mice. The Sel1Lflox/flox mice were then crossed with actin-promoter driven estrogen receptor-Cre fusion protein transgenic (ERCre) mice on the C57BL/6J background (JAX 004781). The last stage of cross generated fl/fl;ERCre+ and the control cohort fl/fl;ERCre- littermates at 1:1 ratio.	To extract RNA from pancreas, mice were anesthetized and pancreas tissues were perfused locally with RNAlater reagent (Qiagen). Tissues were then excised and soaked in RNAlater on ice for stabilization. Total RNA was extracted using Trizol and RNA miniprep kit (Sigma RTN350) with DNaseI digestion (Roche).	Arrays were scanned on an Affymetrix GeneTitan instrument (Affymetrix, Santa Clara, CA).								
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol								
Experimental Factor Name	GENOTYPE														
Experimental Factor Type	genotype														
Comment[SecondaryAccession]	GSE52929														
Comment[GEOReleaseDate]	2013-12-04														
Comment[ArrayExpressSubmissionDate]	2013-12-03														
Comment[GEOLastUpdateDate]	2013-12-05														
Comment[AEExperimentType]	transcription profiling by array														
SDRF File	E-GEOD-52929.sdrf.txt