title: Interaction of the HIV-1 frameshift signal with the ribosome.
abstract: Ribosomal frameshifting on viral RNAs relies on the mechanical properties of structural elements, often pseudoknots and more rarely stem-loops, that are unfolded by the ribosome during translation. In human immunodeficiency virus (HIV)-1 type B a long hairpin containing a three-nucleotide bulge is responsible for efficient frameshifting. This three-nucleotide bulge separates the hairpin in two domains: an unstable lower stem followed by a GC-rich upper stem. Toeprinting and chemical probing assays suggest that a hairpin-like structure is retained when ribosomes, initially bound at the slippery sequence, were allowed multiple EF-G catalyzed translocation cycles. However, while the upper stem remains intact the lower stem readily melts. After the first, and single step of translocation of deacylated tRNA to the 30 S P site, movement of the mRNA stem-loop in the 5' direction is halted, which is consistent with the notion that the downstream secondary structure resists unfolding. Mechanical stretching of the hairpin using optical tweezers only allows clear identification of unfolding of the upper stem at a force of 12.8 +/- 1.0 pN. This suggests that the lower stem is unstable and may indeed readily unfold in the presence of a translocating ribosome.
authors.affiliation: Laboratoire de Chimie et Biologie Structurales, FRC 3115 ICSN-CNRS 1 ave de la terrasse, 91190 Gif-sur-Yvette, France.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Mazauric
authors.forename: Yeonee
authors.initials: Y
authors.lastname: Seol
authors.forename: Satoko
authors.initials: S
authors.lastname: Yoshizawa
authors.forename: Koen
authors.initials: K
authors.lastname: Visscher
authors.forename: Dominique
authors.initials: D
authors.lastname: Fourmy
mesh_terms: D018965:Frameshifting, Ribosomal; D015497:HIV-1 / Q000235:genetics*; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral