PMID- 10559216
OWN - NLM
STAT- MEDLINE
DCOM- 19991214
LR  - 20191210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 47
DP  - 1999 Nov 19
TI  - Assembly of AUF1 oligomers on U-rich RNA targets by sequential dimer association.
PG  - 33374-81
AB  - Many labile mammalian mRNAs are targeted for rapid cytoplasmic turnover by the
      presence of A + U-rich elements (AREs) within their 3'-untranslated regions.
      These elements are selectively recognized by AUF1, a component of a multisubunit 
      complex that may participate in the initiation of mRNA decay. In this study, we
      have investigated the recognition of AREs by AUF1 in vitro using
      oligoribonucleotide substrates. Gel mobility shift assays demonstrated that
      U-rich RNA targets were specifically bound by AUF1, generating two distinct
      RNA-protein complexes in a concentration-dependent manner. Chemical cross-linking
      revealed the interaction of AUF1 dimers to form tetrameric structures involving
      protein-protein interactions in the presence of high affinity RNA targets. From
      these data, a model of AUF1 association with AREs involving sequential dimer
      binding was developed. Using fluorescent RNA substrates, binding parameters of
      AUF1 dimer-ARE and tetramer-ARE equilibria were evaluated in solution by
      fluorescence anisotropy measurements. Using two AUF1 deletion mutants, sequences 
      C-terminal to the RNA recognition motifs are shown to contribute to the formation
      of the AUF1 tetramer.ARE complex but are not obligate for RNA binding activity.
      Kinetic studies demonstrated rapid turnover of AUF1.ARE complexes in solution,
      suggesting that these interactions are very dynamic in character. Taken together,
      these data support a model where ARE-dependent oligomerization of AUF1 may
      function to nucleate the formation of a trans-acting, RNA-destabilizing complex
      in vivo.
FAU - Wilson, G M
AU  - Wilson GM
AD  - Department of Microbiology Wake Forest University School of Medicine,
      Winston-Salem, North Carolina 27157-1064, USA.
FAU - Sun, Y
AU  - Sun Y
FAU - Lu, H
AU  - Lu H
FAU - Brewer, G
AU  - Brewer G
LA  - eng
GR  - CA 52443/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Biopolymers)
RN  - 0 (HNRPD protein, human)
RN  - 0 (Heterogeneous Nuclear Ribonucleoprotein D0)
RN  - 0 (Heterogeneous-Nuclear Ribonucleoprotein D)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Base Sequence
MH  - Binding Sites
MH  - Biopolymers
MH  - Dimerization
MH  - Fluorescence Polarization
MH  - Heterogeneous Nuclear Ribonucleoprotein D0
MH  - *Heterogeneous-Nuclear Ribonucleoprotein D
MH  - Humans
MH  - RNA/*metabolism
MH  - RNA-Binding Proteins/chemistry/*metabolism
MH  - Recombinant Proteins/chemistry/metabolism
EDAT- 1999/11/24 00:00
MHDA- 1999/11/24 00:01
CRDT- 1999/11/24 00:00
PHST- 1999/11/24 00:00 [pubmed]
PHST- 1999/11/24 00:01 [medline]
PHST- 1999/11/24 00:00 [entrez]
AID - 10.1074/jbc.274.47.33374 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Nov 19;274(47):33374-81. doi: 10.1074/jbc.274.47.33374.