Dataset Information

Peptide sequences generated by unexpected splicing

ABSTRACT: Transcripts from the human and mouse CAPN10 cDNA sequences cloned into pcDNA and expressed in cultured cells undergo unexpected splicing. Peptide sequences missing from the spliced version of CAPN10 expressed in COS7 cells were determined by MS analysis.

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

SUBMITTER: Yasuko Ono

PROVIDER: PXD026551 | JPOST Repository | Sat Jan 01 00:00:00 GMT 2022

REPOSITORIES: jPOST

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Publications

Cryptic splicing events result in unexpected protein products from calpain-10 (CAPN10) cDNA.

Ono Yasuko Y Doi Naoko N Shindo Mayumi M Pánico Pablo P Salazar Ana María AM

Biochimica et biophysica acta. Molecular cell research 20211211 3

Calpain-10 (CAPN10) belongs to the calpain superfamily. Genetic polymorphisms of the CAPN10 gene are associated with susceptibility to develop type 2 diabetes mellitus. Although the role of CAPN10 in the pathophysiology of diabetes has been extensively investigated, its biochemical properties are largely unknown. In this report, we made the surprising discovery that CAPN10 cDNA transcripts are subject to cryptic splicing and unexpected protein products were expressed. The same set of splicing pr ...[more]

PMID: 34906616

Similar Datasets

Project description:The splicing regulator PTBP2 controls a program of embryonic splicing required for neuronal maturation. The splicing regulatory proteins PTBP1 and PTBP2 show distinct temporal expression profiles in the developing brain. Neuronal progenitor cells predominantly express PTBP1, whereas developing neurons express high levels of PTBP2, which are subsequently reduced late in neuronal maturation. We show here that PTBP2 and the program of splicing it controls are essential to proper neuronal maturation and survival. To investigate its in vivo function, we generated conditional PTBP2 null alleles in mice. Loss of PTBP2 in neuronal progenitor cells leads to neonatal death without gross defects in brain architecture. Mice with specific depletion of PTBP2 in the cortex and forebrain are viable. However over the first three postnatal weeks, when the normal cortex expands and develops mature circuits, the PTBP2 null cortices degenerate. We find that PTBP2-/- neurons cultured from embryonic brain show the same initial viability as wild type cells with proper early marker expression and neurite outgrowth. Strikingly, between 10 and 20 days in culture PTBP2 null neurons undergo a catastrophic failure to mature and die. To assess the target transcripts leading to these phenotypes, we examined the genomewide splicing changes in the PTBP2 null brains. This identified a large number of mis-regulated exons that share a temporal pattern of regulation; in the absence of PTBP2 many isoforms normally found in adults are precociously expressed in the developing brain. Transcripts following this pattern encode essential neuronal proteins affecting neurite growth, pre- and post-synaptic assembly, and synaptic transmission. Our results define a new genetic regulatory program essential for neuronal survival and maturation, where PTBP2 acts to temporarily repress expression of protein isoforms until the final maturation of the neuron. Mice carrying a conditional floxed PTBP2 allele of PTBP2 were crossed to mice carrying Cre recombinase driven by the nestin promoter. The resulting knockout mutant mouse brains were analyzed for changes in gene expression and alternative splicing. Knockout mice were compared to wildtype littermates. Whole mouse brain polyA plus RNA was isolated from three Nestin-cre knockout embryos at embryonic day 18 and compared to three wildtype littermates. RNA was converted to cDNA and used to probe Affymetrix MJAY splicing sensitive microarrays and analysed by Omniviewer to identify changes in splicing.

			Action	DRS
	CAPN10_20K.mgf	Mgf
	CAPN10_20K.mzid	Mzid
	CAPN10_20K.wiff	Wiff
	CAPN10_20K.wiff.scan	Wiff
	CAPN10_20Kr.mgf	Mgf