TY - JOUR
T1 - An RBD that does not bind RNA
T2 - NMR secondary structure determination and biochemical properties of the C-terminal RNA binding domain from the human U1A protein
AU - Lu, Jirong
AU - Hall, Kathleen B.
N1 - Funding Information:
We thank Dr Changguo Tang for help in implementation of pulse sequences, Professor Dave Cistola for advice and the use of NMR Compass, Mike Hodsdon for his assistance, and Professor Frederick Dahlquist for the E. coli strain (DL39 Avta::Tn5) used for selective labeling. The original human UIA clone was received from Professor W. van Venrooij (Nijmegen); the U1 and U2 snRNA constructs from Professor M. R. Green, and U5 from Professor J. Patton. This work is supported by the Lucille P. Markey Charitable Trust (no. 90-47) (K.B.H. is a Lucille P. Markey Scholar), grant no. IN-36-35 from the American Cancer Society (K.B.H.) and the NIH (postdoctoral fellowship GM16739 to J.L.).
PY - 1995/4/7
Y1 - 1995/4/7
N2 - We have obtained backbone 1H, 15N, and 13C assignments and determined the secondary structure and folding topology of the C-terminal RNA-binding domain (RBD) of the human U1A protein. The secondary structure derived from NOE data is in excellent agreement with the predicted structure from the 1H and 13C chemical shift indices. This 88 amino acid domain exhibits a βαβ-βαβ folding pattern, with conserved RNP1 and RNP2 sequences located in two adjacent strands of a four-strand antiparallel β-sheet. This global folding pattern is typical of this class of RNA binding proteins. Although this domain contains residues that are conserved in all RBDs, its RNA binding properties are very unusual. RNA binding studies show that this domain does not bind U1, U2 or U5 snRNA, an RNA hairpin, rA16, rU16, rC16 or rA3U3GUA4, nor does it show significant association to populations of random sequence RNAs.
AB - We have obtained backbone 1H, 15N, and 13C assignments and determined the secondary structure and folding topology of the C-terminal RNA-binding domain (RBD) of the human U1A protein. The secondary structure derived from NOE data is in excellent agreement with the predicted structure from the 1H and 13C chemical shift indices. This 88 amino acid domain exhibits a βαβ-βαβ folding pattern, with conserved RNP1 and RNP2 sequences located in two adjacent strands of a four-strand antiparallel β-sheet. This global folding pattern is typical of this class of RNA binding proteins. Although this domain contains residues that are conserved in all RBDs, its RNA binding properties are very unusual. RNA binding studies show that this domain does not bind U1, U2 or U5 snRNA, an RNA hairpin, rA16, rU16, rC16 or rA3U3GUA4, nor does it show significant association to populations of random sequence RNAs.
KW - NMR assignments
KW - NMR chemical shift index
KW - RNA binding
KW - U1A RBD(2)
UR - http://www.scopus.com/inward/record.url?scp=0028965970&partnerID=8YFLogxK
U2 - 10.1016/S0022-2836(05)80152-4
DO - 10.1016/S0022-2836(05)80152-4
M3 - Article
C2 - 7723028
AN - SCOPUS:0028965970
VL - 247
SP - 739
EP - 752
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 4
ER -