The neuroprotective WldS gene regulates expression of PTTG1 and erythroid differentiation regulator 1-like gene in mice and human cells.
Wallerian degeneration of injured neuronal axons and synapses is blocked in Wld(S) mutant mice by expression of an nicotinamide mononucleotide adenylyl transferase 1 (Nmnat-1)/truncated-Ube4b chimeric gene. The protein product of the Wld(S) gene localizes to neuronal nuclei. Here we show that Wld(S) protein expression selectively alters mRNA levels of other genes in Wld(S) mouse cerebellum in vivo and following transfection of human embryonic kidney (HEK293) cells in vitro. The largest changes, identified by microarray analysis and quantitative real-time polymerase chain reaction of cerebellar mRNA, were an approximate 10-fold down-regulation of pituitary tumour-transforming gene-1 (pttg1) and an approximate 5-fold up-regulation of a structural homologue of erythroid differentiation regulator-1 (edr1l-EST). Transfection of HEK293 cells with a Wld(S)-eGFP construct produced similar changes in mRNA levels for these and seven other genes, suggesting that regulation of gene expression by Wld(S) is conserved across different species, including humans. Similar modifications in mRNA levels were mimicked for some of the genes (including pttg1) by 1 mm nicotinamide adenine dinucleotide (NAD). However, expression levels of most other genes (including edr1l-EST) were insensitive to NAD. Pttg1(-/-) mutant mice showed no neuroprotective phenotype. Transfection of HEK293 cells with constructs comprising either full-length Nmnat-1 or the truncated Ube4b fragment (N70-Ube4b) demonstrated selective effects of Nmnat-1 (down-regulated pttg1) and N70-Ube4b (up-regulated edr1l-EST) on mRNA levels. Similar changes in pttg1 and edr1l-EST were observed in the mouse NSC34 motor neuron-like cell line following stable transfection with Wld(S). Together, the data suggest that the Wld(S) protein co-regulates expression of a consistent subset of genes in both mouse neurons and human cells. Targeting Wld(S)-induced gene expression may lead to novel therapies for neurodegeneration induced by trauma or by disease in humans.
Gillingwater
Thomas H
TH
Centre for Neuroscience Research, University of Edinburgh, 1 George Square, Edinburgh EH8 9JZ, UK. t.gillingwater@ed.ac.uk
Wishart
Thomas M
TM
Chen
Philip E
PE
Haley
Jane E
JE
Robertson
Kevin
K
MacDonald
Stephen H-F
SH
Middleton
Susan
S
Wawrowski
Kolja
K
Shipston
Michael J
MJ
Melmed
Shlomo
S
Wyllie
David J A
DJ
Skehel
Paul A
PA
Coleman
Michael P
MP
Ribchester
Richard R
RR
D000818:Animals; D002460:Cell Line; D005786:Gene Expression Regulation / Q000235:genetics; D018390:Gene Targeting; D015316:Genetic Therapy; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D064249:Securin; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases / Q000096:biosynthesis* / Q000235:genetics; D014855:Wallerian Degeneration / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D014947:Wounds and Injuries / Q000235:genetics / Q000378:metabolism / Q000628:therapy
D008565:Membrane Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D064249:Securin; D025521:Tumor Suppressor Proteins; C442580:Wld protein, mouse; C505963:erythroid differentiation regulator 1, mouse; C578002:pituitary tumor-transforming protein 1, human; D043743:Ubiquitin-Protein Ligase Complexes; C473697:UBE4B protein, human; C471292:Ube4b protein, mouse; D044767:Ubiquitin-Protein Ligases; D009612:Nicotinamide-Nucleotide Adenylyltransferase; C510972:Nmnat1 protein, mouse