Identification of regulatory cis-acting elements for alternative splicing of presenilin 2 exon 5 under hypoxic stress conditions.
An alternatively spliced form of the presenilin 2 (PS2) gene lacking exon 5 (PS2V) was found in human brains with sporadic Alzheimer's disease. PS2V was induced by hypoxic stress in human neuroblastoma SK-N-SH cells, indicating that hypoxic stress affects the splicing machineries for PS2 exon 5. Here, we identified the critical cis-acting element (sec 2) on the PS2 pre-mRNA responsible for the aberrant splicing of PS2 exon 5 under hypoxic stress conditions. The element was composed of 23 nucleotides in exon 5 and RNA structural analyses showed a stem-loop structure in this sequence. Treatment with an antisense oligonucleotide directed toward the cis-acting element caused an increase in exon 5 inclusion. These results indicate that the sec 2 identified in this study is a novel regulatory element for exon 5 splicing under stress conditions and that trans-acting factors could specifically bind to the element to skip exon 5 of PS2.
Higashide
Satoshi
S
Division of Structural Cellular Biology, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Morikawa
Keisuke
K
Okumura
Masayo
M
Kondo
Shinichi
S
Ogata
Maiko
M
Murakami
Tomohiko
T
Yamashita
Akinori
A
Kanemoto
Soshi
S
Manabe
Takayuki
T
Imaizumi
Kazunori
K
D017398:Alternative Splicing; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D000860:Hypoxia / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D008565:Membrane Proteins / Q000235:genetics*; D008958:Models, Molecular; D016296:Mutagenesis; D009447:Neuroblastoma; D009532:Nickel; D009584:Nitrogen; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology; D053766:Presenilin-2; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013312:Stress, Physiological / Q000235:genetics / Q000378:metabolism; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D020320:Oligoribonucleotides, Antisense; C508861:PSEN2 protein, human; D053766:Presenilin-2; D012333:RNA, Messenger; D016875:tau Proteins; C003320:sodium nitride; C022838:nickel chloride; D009532:Nickel; D009584:Nitrogen