RT Journal Article
SR Electronic
T1 Evidence of an Unusual Poly(A) RNA Signature Detected by High-throughput Chemical Mapping
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 281147
DO 10.1101/281147
A1 Roger Wellington-Oguri
A1 Eli Fisker
A1 Mathew Zada
A1 Michelle Wiley
A1 Eterna Players
YR 2018
UL http://biorxiv.org/content/early/2018/04/30/281147.abstract
AB Homopolymeric adenosine RNA plays numerous roles in both cells and non-cellular genetic material, and for lack of evidence to the contrary, it is generally accepted to form a random coil under physiological conditions. However, chemical mapping data generated by the Eterna Massive Open Laboratory indicates that a poly (A) sequence of length seven or more, at pH 8.0 and MgCl concentrations of 10 mM, develops unexpected protection to selective 2’-hydroxyl acylation read out by primer extension (SHAPE) and dimethyl sulfate (DMS) chemical probing. This protection first appears in poly(A) sequences of length 7 and grows to its maximum strength at length ~10. In a long poly(A) sequence, substitution of a single A by any other nucleotide disrupts the protection, but only for the 6 or so nucleotides on the 5 ‘ side of the substitution.SHAPE, Selective 2 hydroxyl acylation analyzed by primer extension;NMIA, N-methylisatoic anhydride;1M7, 1-methyl-7-nitroisatoic anhydride;DMS, Dimethyl sulfide,PABPN1, Nuclear poly(A) binding protein 1;PAP, poly(A) polymerase;RRM, RNA recognition motif.