the 5′ leader of the mrna encoding the mouse neurotrophin receptor trkb contains two internal ribosomal entry sites that are differentially regulated信使rna编码的5u2032领导人鼠标生成受体trkb包含两个不同的内部核糖体进入位点监管.pdfVIP

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the 5′ leader of the mrna encoding the mouse neurotrophin receptor trkb contains two internal ribosomal entry sites that are differentially regulated信使rna编码的5u2032领导人鼠标生成受体trkb包含两个不同的内部核糖体进入位点监管.pdf

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the 5′ leader of the mrna encoding the mouse neurotrophin receptor trkb contains two internal ribosomal entry sites that are differentially regulated信使rna编码的5u2032领导人鼠标生成受体trkb包含两个不同的内部核糖体进入位点监管

The 5 9 Leader of the mRNA Encoding the Mouse Neurotrophin Receptor TrkB Contains Two Internal Ribosomal Entry Sites that Are Differentially Regulated 1 1¤ 1 1,2 Stephanie L. Timmerman , Jennifer S. Pfingsten , Jeffrey S. Kieft , Les A. Krushel * 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver School of Medicine, Aurora, Colorado, United States of America, 2 Department of Pharmacology, University of Colorado Denver School of Medicine, Aurora, Colorado, United States of America Abstract A single internal ribosomal entry site (IRES) in conjunction with IRES transactivating factors (ITAFs) is sufficient to recruit the translational machinery to a eukaryotic mRNA independent of the cap structure. However, we demonstrate that the mouse TrkB mRNA contains two independent IRESes. The mouse TrkB mRNA consists of one of two 5 9 leaders (1428 nt and 448 nt), both of which include the common 39 exon (Ex2, 344 nt). Dicistronic RNA transfections and in vitro translation of monocistronic RNA demonstrated that both full-length 59 leaders, as well as Ex2, exhibit IRES activity indicating the IRES is located within Ex2. Additional analysis of the upstream sequences demonstrated that the first 260 nt of exon 1 (Ex1a) also contains an IRES. Dicistronic RNA transfections into SH-SY5Y cells showed the Ex1a IRES is constitutively active. However, the Ex2 IRES is only active in response to retinoic acid induced neural differentiation, a state which correlates with the synthesis of the ITAF polypyrimidine tract binding protein (PTB1). Correspondingly, addition or knock-down of PTB1 altered Ex2, but not Ex1a IRES activity in vitro and ex vivo, respectively. These results demonst