PMID- 10710437
OWN - NLM
STAT- MEDLINE
DCOM- 20000331
LR  - 20190501
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 28
IP  - 7
DP  - 2000 Apr 1
TI  - RNA-binding analyses of HuC and HuD with the VEGF and c-myc 3'-untranslated 
      regions using a novel ELISA-based assay.
PG  - E20
AB  - Human members of the ELAV family, referred to as ELAV-like proteins (ELPs), 
      include HuC, HuD, Hel-N1 and HuR. These proteins bind to AU-rich elements in the 
      3'-untranslated regions (3'-UTRs) of many growth-related mRNAs, including c-myc 
      and VEGF, and may participate in regulating the stability of these transcripts. 
      Here, I have developed an enzyme-linked immunosorbent assay (ELISA) which can 
      rapidly assess the RNA-protein-binding properties of ELPs. With this assay, I 
      demonstrate that HuC and HuD bind to the VEGF 3'-UTR regulatory segment (VRS) and 
      to the c- myc 3'-UTR in a specific and concentration-dependent pattern, with both 
      proteins showing a greater affinity for the VRS. Further analysis of the VRS 
      indicated that the binding affinity was greater for the 3'-end where the majority 
      of AU motifs reside. Binding to the VRS could be competed by both proteins as 
      well as a poly(U) ribohomopolymer. The binding could not be competed by other 
      ribohomopolymers or serum from patients with high titer anti-HuD antibodies. In 
      summary, this assay provides a rapid analysis of ELP-RNA binding which can be 
      utilized for further characterization of RNA-binding properties and for 
      identification of competitor molecules for in vivo functional analysis of ELPs.
FAU - King, P H
AU  - King PH
AD  - Department of Neurology, University of Alabama, 1235 Jefferson Tower, 625 South 
      19th Street, Birmingham, AL 35294-0007, USA. pking@uab.edu
LA  - eng
GR  - P30 CA013148/CA/NCI NIH HHS/United States
GR  - 5P50 CA13148/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (3' Untranslated Regions)
RN  - 0 (Antibodies)
RN  - 0 (ELAV Proteins)
RN  - 0 (ELAV-Like Protein 3)
RN  - 0 (ELAV-Like Protein 4)
RN  - 0 (ELAVL3 protein, human)
RN  - 0 (ELAVL4 protein, human)
RN  - 0 (Endothelial Growth Factors)
RN  - 0 (Lymphokines)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Proto-Oncogene Proteins c-myc)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Vascular Endothelial Growth Factor A)
RN  - 0 (Vascular Endothelial Growth Factors)
RN  - 27416-86-0 (Poly U)
SB  - IM
MH  - 3' Untranslated Regions/*metabolism
MH  - Antibodies/immunology
MH  - Base Sequence
MH  - Binding, Competitive
MH  - ELAV Proteins
MH  - ELAV-Like Protein 3
MH  - ELAV-Like Protein 4
MH  - Endothelial Growth Factors/*genetics
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Humans
MH  - Lymphokines/*genetics
MH  - Molecular Sequence Data
MH  - Nerve Tissue Proteins/genetics/immunology/*metabolism
MH  - Paraneoplastic Syndromes, Nervous System/immunology
MH  - Poly U/metabolism
MH  - Protein Binding
MH  - Proto-Oncogene Proteins c-myc/*genetics
MH  - RNA-Binding Proteins/genetics/immunology/*metabolism
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Regulatory Sequences, Nucleic Acid
MH  - Tumor Cells, Cultured
MH  - Vascular Endothelial Growth Factor A
MH  - Vascular Endothelial Growth Factors
PMC - PMC102803
EDAT- 2000/03/10 00:00
MHDA- 2000/03/10 00:01
PMCR- 2000/04/01
CRDT- 2000/03/10 00:00
PHST- 2000/03/10 00:00 [pubmed]
PHST- 2000/03/10 00:01 [medline]
PHST- 2000/03/10 00:00 [entrez]
PHST- 2000/04/01 00:00 [pmc-release]
AID - gnd020 [pii]
AID - 10.1093/nar/28.7.e20 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2000 Apr 1;28(7):E20. doi: 10.1093/nar/28.7.e20.