Passa al contenuto
Merck

An RNA binding protein promotes axonal integrity in peripheral neurons by destabilizing REST.

The Journal of neuroscience : the official journal of the Society for Neuroscience (2014-12-17)
Francesca Cargnin, Tamilla Nechiporuk, Karin Müllendorff, Deborah J Stumpo, Perry J Blackshear, Nurit Ballas, Gail Mandel
ABSTRACT

The RE1 Silencing Transcription Factor (REST) acts as a governor of the mature neuronal phenotype by repressing a large consortium of neuronal genes in non-neuronal cells. In the developing nervous system, REST is present in progenitors and downregulated at terminal differentiation to promote acquisition of mature neuronal phenotypes. Paradoxically, REST is still detected in some regions of the adult nervous system, but how REST levels are regulated, and whether REST can still repress neuronal genes, is not known. Here, we report that homeostatic levels of REST are maintained in mature peripheral neurons by a constitutive post-transcriptional mechanism. Specifically, using a three-hybrid genetic screen, we identify the RNA binding protein, ZFP36L2, associated previously only with female fertility and hematopoiesis, and show that it regulates REST mRNA stability. Dorsal root ganglia in Zfp36l2 knock-out mice, or wild-type ganglia expressing ZFP36L2 shRNA, show higher steady-state levels of Rest mRNA and protein, and extend thin and disintegrating axons. This phenotype is due, at least in part, to abnormally elevated REST levels in the ganglia because the axonal phenotype is attenuated by acute knockdown of REST in Zfp36l2 KO DRG explants. The higher REST levels result in lower levels of target genes, indicating that REST can still fine-tune gene expression through repression. Thus, REST levels are titrated in mature peripheral neurons, in part through a ZFP36L2-mediated post-transcriptional mechanism, with consequences for axonal integrity.

MATERIALI
N° Catalogo
Marchio
Descrizione del prodotto

Sigma-Aldrich
Acetone, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetone, suitable for HPLC, ≥99.9%
Sigma-Aldrich
Acetone, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
Sigma-Aldrich
Saccarosio, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Saccarosio, ≥99.5% (GC)
Sigma-Aldrich
Sodio cloruro, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Cloruro di magnesio, for molecular biology, 1.00 M±0.01 M
Sigma-Aldrich
Cloruro di magnesio, anhydrous, ≥98%
Sigma-Aldrich
Saccarosio, ≥99.5% (GC), BioXtra
Sigma-Aldrich
Saccarosio, BioUltra, for molecular biology, ≥99.5% (HPLC)
Sigma-Aldrich
Sodio cloruro, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Sodio cloruro, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Sodio cloruro, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
USP
Saccarosio, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Streptomicina, powder, BioReagent, suitable for cell culture
SAFC
Sodio cloruro, 5 M
Sigma-Aldrich
Selenium, powder, −100 mesh, 99.99% trace metals basis
Sigma-Aldrich
Saccarosio, ≥99.5% (GC), BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Acetone, ACS reagent, ≥99.5%
Sigma-Aldrich
Saccarosio, ≥99.5% (GC)
Sigma-Aldrich
Cloruro di magnesio, powder, <200 μm
Sigma-Aldrich
Selenium, powder, −100 mesh, ≥99.5% trace metals basis
Sigma-Aldrich
Sodio cloruro, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
Saccarosio, ≥99.5% (GC), Grade II, suitable for plant cell culture
Sigma-Aldrich
Sodio cloruro, 99.999% trace metals basis
USP
Acetone, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Sodio cloruro, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
Cloruro di magnesio, BioUltra, for molecular biology, 2 M in H2O
Sigma-Aldrich
Acetone, histological grade, ≥99.5%
Sigma-Aldrich
Cloruro di magnesio, BioReagent, suitable for insect cell culture, ≥97.0%