H-1, C-13, AND N-15 NMR ASSIGNMENTS AND GLOBAL FOLDING PATTERN OF THE RNA-BINDING DOMAIN OF THE HUMAN HNRNP C-PROTEINS

被引:141
作者
WITTEKIND, M
GORLACH, M
FRIEDRICHS, M
DREYFUSS, G
MUELLER, L
机构
[1] UNIV PENN,SCH MED,HOWARD HUGHES MED INST,PHILADELPHIA,PA 19104
[2] UNIV PENN,SCH MED,DEPT BIOCHEM & BIOPHYS,PHILADELPHIA,PA 19104
关键词
D O I
10.1021/bi00142a013
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The hnRNP C1 and C2 proteins are abundant nuclear proteins that bind avidly to heterogeneous nuclear RNAs (hnRNAs) and appear to be involved with pre-mRNA processing. The RNA-binding activity of the hnRNP C proteins is contained in the amino-terminal 94 amino acid RNA-binding domain (RBD) that is identical for these two proteins. We have obtained the H-1, C-13, and N-15 NMR assignments for the RBD of the human hnRNP C proteins. The assignment process was facilitated by extensive utilization of three- and four-dimensional heteronuclear-edited spectra. Sequential assignments of the backbone resonances were made using a combination of N-15-edited 3D NOESY-HMQC, 3D TOCSY-HMQC, and 3D TOCSY-NOESY-HSQC as well as 3D HNCA, HNCO, and HCACO spectra. Side-chain resonances were assigned using 3D HCCH-COSY and 3D HCH-TOCSY spectra. Four-dimensional C-13/C-13-edited NOESY and C-13/N-15-edited NOESY experiments were used to unambigously resolve NOEs. The overall global folding pattern was established by calculating a set of preliminary structures using constraints derived from the sequential NOEs and a small number of long-range NOEs. The beta-alpha-beta-beta-alpha-beta-domain structure exhibits an antiparallel beta-sheet with the conserved RNP 1 and RNP 2 sequences [Dreyfuss et al. (1988) Trends Biochem. Sci. 13, 86-91] located adjacent to one another as the two inner strands of the beta-sheet.
引用
收藏
页码:6254 / 6265
页数:12
相关论文
共 76 条
[1]   MESSENGER-RNA POLYADENYLATE-BINDING PROTEIN - GENE ISOLATION AND SEQUENCING AND IDENTIFICATION OF A RIBONUCLEOPROTEIN CONSENSUS SEQUENCE [J].
ADAM, SA ;
NAKAGAWA, T ;
SWANSON, MS ;
WOODRUFF, TK ;
DREYFUSS, G .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (08) :2932-2943
[2]   RNA-BINDING PROTEINS AS DEVELOPMENTAL REGULATORS [J].
BANDZIULIS, RJ ;
SWANSON, MS ;
DREYFUSS, G .
GENES & DEVELOPMENT, 1989, 3 (04) :431-437
[3]   MLEV-17-BASED TWO-DIMENSIONAL HOMONUCLEAR MAGNETIZATION TRANSFER SPECTROSCOPY [J].
BAX, A ;
DAVIS, DG .
JOURNAL OF MAGNETIC RESONANCE, 1985, 65 (02) :355-360
[4]   PRACTICAL ASPECTS OF PROTON CARBON CARBON PROTON 3-DIMENSIONAL CORRELATION SPECTROSCOPY OF C-13-LABELED PROTEINS [J].
BAX, A ;
CLORE, GM ;
DRISCOLL, PC ;
GRONENBORN, AM ;
IKURA, M ;
KAY, LE .
JOURNAL OF MAGNETIC RESONANCE, 1990, 87 (03) :620-627
[5]   NATURAL ABUNDANCE N-15 NMR BY ENHANCED HETERONUCLEAR SPECTROSCOPY [J].
BODENHAUSEN, G ;
RUBEN, DJ .
CHEMICAL PHYSICS LETTERS, 1980, 69 (01) :185-189
[6]   TOWARD COMPLETE H-1-NMR SPECTRA IN PROTEINS [J].
BROWN, SC ;
WEBER, PL ;
MUELLER, L .
JOURNAL OF MAGNETIC RESONANCE, 1988, 77 (01) :166-169
[7]  
BRUNGER AT, 1990, XPLOR VERSION 2 1 US
[8]   PRIMARY STRUCTURES OF THE HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN A2-PROTEIN, B1-PROTEIN, AND C2-PROTEIN - A DIVERSITY OF RNA-BINDING PROTEINS IS GENERATED BY SMALL PEPTIDE INSERTS [J].
BURD, CG ;
SWANSON, MS ;
GORLACH, M ;
DREYFUSS, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (24) :9788-9792
[9]  
BURD CG, 1991, MOL CELL BIOL, V7, P3419
[10]   HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEINS - ROLE IN RNA SPLICING [J].
CHOI, YD ;
GRABOWSKI, PJ ;
SHARP, PA ;
DREYFUSS, G .
SCIENCE, 1986, 231 (4745) :1534-1539