Patterning and gastrulation defects caused by the t(w18) lethal are due to loss of Ppp2r1a.

Biol Open. 2017 Jun 15;6(6):752-764

Lisette Lange ¹ , Matthias Marks ² , Jinhua Liu ² , Lars Wittler ² , Hermann Bauer ² ,

Lisette Lange ¹ , Matthias Marks ² , Jinhua Liu ² , Lars Wittler ² , Hermann Bauer ² , Sandra Piehl ² , Gabriele Bläß ² , Bernd Timmermann ³ , Bernhard G Herrmann ⁴

+ et al

Author information

¹ Free University Berlin, Department of Biology, Chemistry and Pharmacy, Takustrasse 3, Berlin 14195, Germany.
² Max Planck Institute for Molecular Genetics, Department Developmental Genetics, Ihnestraße 63-73, Berlin 14195, Germany.
³ Max Planck Institute for Molecular Genetics, Sequencing Core Facility, Ihnestraße 63-73, Berlin 14195, Germany.
⁴ Charité-University Medicine Berlin, Institute for Medical Genetics, Campus Benjamin Franklin, Hindenburgdamm 30, Berlin 12203, Germany.

摘要

The mouse t haplotype, a variant 20âcM genomic region on Chromosome 17, harbors 16 embryonic control genes identified by recessive lethal mutations isolated from wild mouse populations. Due to technical constraints so far only one of these, the t(w5) lethal, has been cloned and molecularly characterized. Here we report the molecular isolation of the t(w18) lethal. Embryos carrying the t(w18) lethal die from major gastrulation defects commencing with primitive streak formation at E6.5. We have used transcriptome and marker gene analyses to describe the molecular etiology of the t(w18) phenotype. We show that both WNT and Nodal signal transduction are impaired in the mutant epiblast, causing embryonic patterning defects and failure of primitive streak and mesoderm formation. By using a candidate gene approach, gene knockout by homologous recombination and genetic rescue, we have identified the gene causing the t(w18) phenotype as Ppp2r1a, encoding the PP2A scaffolding subunit PR65alpha. Our work highlights the importance of phosphatase 2A in embryonic patterning, primitive streak formation, gastrulation, and mesoderm formation downstream of WNT and Nodal signaling.

KEYWORDS: Mesoderm, PP2A phosphatase, Primitive streak, Signal transduction, t haplotype, tw18

原始数据

保存测序数据

Sample name	Organism	Experiment title	Sample type	Library instrument	Attributes
Sample name	Organism	Experiment title	Sample type	Library instrument	development stage	genotype	source name	strain	tissue
tw18 /tw18	Mus musculus	GSM2387400: tw18 /tw18; Mus musculus; RNA-Seq	RNA-Seq	Illumina HiSeq 2000	E6.5	tw18/tw18	tw18 homozygote epiblast	129/SvEv	epiblast
tw18/+	Mus musculus	GSM2387399: tw18/+; Mus musculus; RNA-Seq	RNA-Seq	Illumina HiSeq 2000	E6.5	tw18/+	tw18 heterozygote epiblast	129/SvEv	epiblast
WT	Mus musculus	GSM2387398: WT; Mus musculus; RNA-Seq	RNA-Seq	Illumina HiSeq 2000	E6.5	wildtype	wild type epiblast	129/SvEv	epiblast