Alteration of POLDIP3 splicing associated with loss of function of TDP-43 in tissues affected with ALS.
Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease caused by selective loss of motor neurons. In the ALS motor neurons, TAR DNA-binding protein of 43 kDa (TDP-43) is dislocated from the nucleus to cytoplasm and forms inclusions, suggesting that loss of a nuclear function of TDP-43 may underlie the pathogenesis of ALS. TDP-43 functions in RNA metabolism include regulation of transcription, mRNA stability, and alternative splicing of pre-mRNA. However, a function of TDP-43 in tissue affected with ALS has not been elucidated. We sought to identify the molecular indicators reflecting on a TDP-43 function. Using exon array analysis, we observed a remarkable alteration of splicing in the polymerase delta interacting protein 3 (POLDIP3) as a result of the depletion of TDP-43 expression in two types of cultured cells. In the cells treated with TDP-43 siRNA, wild-type POLDIP3 (variant-1) decreased and POLDIP3 lacking exon 3 (variant-2) increased. The RNA binding ability of TDP-43 was necessary for inclusion of POLDIP3 exon 3. Moreover, we found an increment of POLDIP3 variant-2 mRNA in motor cortex, spinal cord and spinal motor neurons collected by laser capture microdissection with ALS. Our results suggest a loss of TDP-43 function in tissues affected with ALS, supporting the hypothesis that a loss of function of TDP-43 underlies the pathogenesis of ALS.
Department of Pathology, Brain Research Institute, Niigata University, Niigata, Japan.
Atsushi
A
Shiga
Tomohiko
T
Ishihara
Akinori
A
Miyashita
Misaki
M
Kuwabara
Taisuke
T
Kato
Norihiro
N
Watanabe
Akie
A
Yamahira
Chigusa
C
Kondo
Akio
A
Yokoseki
Masuhiro
M
Takahashi
Ryozo
R
Kuwano
Akiyoshi
A
Kakita
Masatoyo
M
Nishizawa
Hitoshi
H
Takahashi
Osamu
O
Onodera
D017398:Alternative Splicing; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D048429:Cell Size; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005091:Exons; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D020033:Protein Isoforms; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C495327:POLDIP3 protein, human; D020033:Protein Isoforms; D012322:RNA Precursors; D016601:RNA-Binding Proteins