Solution structure of the second RNA recognition motif (RRM) domain of murine T cell intracellular antigen-1 (TIA-1) and its RNA recognition mode.
T cell intracellular antigen-1 (TIA-1), an apoptosis promoting factor, functions as a splicing regulator for the Fas pre-mRNA. TIA-1 possesses three RNA recognition motifs (RRMs) and a glutamine-rich domain. The second RRM (RRM2) is necessary and sufficient for tight, sequence-specific binding to the uridine-rich sequences buried around the 5' splice sites. In the present study, we solved the solution structure of the murine TIA-1 RRM2 by heteronuclear-nuclear magnetic resonance spectroscopy. The TIA-1 RRM2 adopts the RRM fold (betaalphabetabetaalphabeta) and possesses an extra beta-strand between beta2 and beta3, which forms an additional beta-sheet with the C-terminal part of beta2. We refer to this structure as the beta2-beta2' beta-loop. Interestingly, this characteristic beta-loop structure is conserved among a number of RRMs, including the U2AF65 RRM2 and the Sex-lethal RRM1 and RRM2, which also bind to uridine-rich RNAs. Furthermore, we identified a new sequence motif in the beta2-beta2' beta-loop, the DxxT motif. Chemical shift perturbation analyses of both the main and side chains upon binding to the uridine pentamer RNA revealed that most of the beta-sheet surface, including the beta2-beta2' beta-loop, is involved in the RNA binding. An investigation of the chemical shift perturbation revealed similarity in the RNA recognition modes between the TIA-1 and U2AF65 RRMs.
Kuwasako
Kanako
K
Protein Research Group, Genomic Sciences Center, Yokohama Institute, RIKEN, Tsurumi-ku, Yokohama 230-0045, Japan.
Takahashi
Mari
M
Tochio
Naoya
N
Abe
Chikage
C
Tsuda
Kengo
K
Inoue
Makoto
M
Terada
Takaho
T
Shirouzu
Mikako
M
Kobayashi
Naohiro
N
Kigawa
Takanori
T
Taguchi
Seiichi
S
Tanaka
Akiko
A
Hayashizaki
Yoshihide
Y
Güntert
Peter
P
Muto
Yutaka
Y
Yokoyama
Shigeyuki
S
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology; D012261:Ribonucleoproteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D012996:Solutions; D000072668:Splicing Factor U2AF; D013499:Surface Properties; D000074019:T-Cell Intracellular Antigen-1
D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D012996:Solutions; D000072668:Splicing Factor U2AF; D000074019:T-Cell Intracellular Antigen-1; C499367:Tia1 protein, mouse; C000607494:U2AF2 protein, human; C000607495:Zrsr1 protein, mouse