The Elav-like proteins bind to a conserved regulatory element in the 3'-untranslated region of GAP-43 mRNA.
Previous studies have identified three brain proteins (40, 65 and 95 kDa, respectively) that specifically bind to the 3'-untranslated region of GAP-43 mRNA. In this study, using a specific monoclonal antibody, we now show that the 40-kDa proteins are members of the Elav-like protein family. This family of specific RNA-binding proteins comprise three neural specific members called HuD, HuC, and Hel-N1. We have shown that purified recombinant HuD can bind with high affinity to GAP-43 mRNA. In addition, we have mapped the binding site to a highly conserved 26-nucleotide sequence within the regulatory element. The binding of HuD to this site is readily displaced by RNA oligonucleotides encoding other HuD binding sites. We also show that only the first and second RNA binding domains of HuD are required for selective binding to GAP-43 mRNA.
Chung
S
S
Program in Molecular Pharmacology and Therapeutics, Memorial Sloan Kettering Cancer Center, New York, New York 10021, USA.
Eckrich
M
M
Perrone-Bizzozero
N
N
Kohn
D T
DT
Furneaux
H
H
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D051959:ELAV Proteins; D000067777:ELAV-Like Protein 2; D019922:GAP-43 Protein; D005786:Gene Expression Regulation; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins; D012045:Regulatory Sequences, Nucleic Acid
D051959:ELAV Proteins; D000067777:ELAV-Like Protein 2; C490170:Elavl2 protein, rat; D019922:GAP-43 Protein; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins