Tethering KSRP, a decay-promoting AU-rich element-binding protein, to mRNAs elicits mRNA decay.
Inherently unstable mRNAs contain AU-rich elements (AREs) in their 3' untranslated regions that act as mRNA stability determinants by interacting with ARE-binding proteins (ARE-BPs). We have destabilized two mRNAs by fusing sequence-specific RNA-binding proteins to KSRP, a decay-promoting ARE-BP, in a tethering assay. These results support a model that KSRP recruits mRNA decay machinery/factors to elicit decay. The ability of tethered KSRP to elicit mRNA decay depends on functions of known mRNA decay enzymes. By targeting the Rev response element of human immunodeficiency virus type 1 by using Rev-KSRP fusion protein, we degraded viral mRNA, resulting in a dramatic reduction of viral replication. These results provide a foundation for the development of novel therapeutic strategies to inhibit specific gene expression in patients with acquired or hereditary diseases.
Chou
Chu-Fang
CF
Department of Biochemistry & Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Mulky
Alok
A
Maitra
Sushmit
S
Lin
Wei-Jye
WJ
Gherzi
Roberto
R
Kappes
John
J
Chen
Ching-Yi
CY
D020413:3' Untranslated Regions; D020816:Amino Acid Motifs; D001665:Binding Sites; D015152:Blotting, Northern; D017930:Genes, Reporter; D005914:Globins / Q000235:genetics; D006207:Half-Life; D006367:HeLa Cells; D006801:Humans; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D020871:RNA Stability / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D020413:3' Untranslated Regions; C105789:KHSRP protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D005914:Globins