Binding of U1A protein to the 3' untranslated region of its pre-mRNA

TY - JOUR

T1 - Binding of U1A protein to the 3' untranslated region of its pre-mRNA

AU - Grainger, Richard J.

AU - Norman, David G.

AU - Lilley, David M.J.

N1 - Funding Information: We thank Dr G. Varani and colleagues for supplying U1A protein, and for valuable discussion. We thank Frank Walter for discussion, and the Cancer Research Campaign and the BBSRC for generous financial support.

PY - 1999/5/14

Y1 - 1999/5/14

N2 - We have studied the global structure of the U1A 3' untranslated region (UTR) element using fluorescence resonance energy transfer. Comparison of a single UTR-box with a series of oligoadenine bulges indicates that the UTR-box introduces a significant kink into the axis of the RNA, and quantification of the results suggests an included bend angle of approximately 100°(i.e. 80°from linear). The complete 3' UTR element is also severely kinked by the two UTR-boxes. We can observe binding of U1A protein to the 3'-UTR element by a change in the fluorescence anisotropy of Cy3 attached to one of the helical ends. In parallel with the binding, we observe a marked increase in fluoresence resonance energy transfer efficiency between fluorophores attached at the two 5' termini, indicating a significant change in global conformation induced by the binding of the protein.

AB - We have studied the global structure of the U1A 3' untranslated region (UTR) element using fluorescence resonance energy transfer. Comparison of a single UTR-box with a series of oligoadenine bulges indicates that the UTR-box introduces a significant kink into the axis of the RNA, and quantification of the results suggests an included bend angle of approximately 100°(i.e. 80°from linear). The complete 3' UTR element is also severely kinked by the two UTR-boxes. We can observe binding of U1A protein to the 3'-UTR element by a change in the fluorescence anisotropy of Cy3 attached to one of the helical ends. In parallel with the binding, we observe a marked increase in fluoresence resonance energy transfer efficiency between fluorophores attached at the two 5' termini, indicating a significant change in global conformation induced by the binding of the protein.

KW - Fluorescence resonance energy transfer

KW - RNA splicing

KW - RNA-protein interaction

UR - https://www.scopus.com/pages/publications/0033043229

U2 - 10.1006/jmbi.1999.2717

DO - 10.1006/jmbi.1999.2717

M3 - Article

C2 - 10329165

AN - SCOPUS:0033043229

SN - 0022-2836

VL - 288

SP - 585

EP - 594

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 4

ER -