NMR signal assignment of the polyuridine tract of the single-stranded RNA complexed with Sxl RBD1-RBD2 by using residue selective [5-(2)H]uridine substitutions.
Using [5-(2)H]uridine phosphoramidite, we synthesized a series of 2H-labeled Drosophila Sex-lethal (Sxl) target RNAs, in which all the uridine residues except one were specifically replaced by [5-(2)H]uridine. By observing the H5-H6 cross peaks of RNA in the TOCSY spectra, we unambiguously assigned all the base proton resonances of the target RNA in a Sxl x RNA complex. Furthermore, it was shown that Sxl differently recognizes A and G in a position prior to the polyuridine tract.
Kim
I
I
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan.
Muto
Y
Y
Hosono
K
K
Kitamura
A
A
Watanabe
S
S
Ohtsuki
T
T
Kawai
G
G
Takaku
H
H
Watanabe
K
K
Sakamoto
H
H
Shimura
Y
Y
Yokoyama
S
S
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003903:Deuterium; D004330:Drosophila / Q000235:genetics; D029721:Drosophila Proteins; D007301:Insect Hormones / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D011072:Poly U; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics*; D014529:Uridine
D029721:Drosophila Proteins; D007301:Insect Hormones; D016601:RNA-Binding Proteins; C067777:Sxl protein, Drosophila; D011072:Poly U; D012313:RNA; D003903:Deuterium; D014529:Uridine