Shine dalgarno sequence6/10/2023 108090000623 proteins and genes Proteins 0.000 claims abstract description 43.102000004169 proteins and genes Human genes 0.000 claims abstract description 43.Publication of US20050153339A1 publication Critical patent/US20050153339A1/en Priority to US11/447,892 priority patent/US7405059B2/en Application granted granted Critical Publication of US7094573B2 publication Critical patent/US7094573B2/en Priority to US12/142,313 priority patent/US20080274503A1/en Status Expired - Fee Related legal-status Critical Current Anticipated expiration legal-status Critical Links ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.) Filing date Publication date Priority claimed from US39143302P external-priority Application filed by Human Genome Sciences Inc filed Critical Human Genome Sciences Inc Priority to US11/004,853 priority Critical patent/US7094573B2/en Assigned to HUMAN GENOME SCIENCES, INC. Original Assignee Human Genome Sciences Inc Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.) ( en Inventor Michael Laird Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) Granted Application number US11/004,853 Other versions US7094573B2 Google Patents Modified shine-dalgarno sequences and methods of use thereofĭownload PDF Info Publication number US20050153339A1 US20050153339A1 US11/004,853 US485304A US2005153339A1 US 20050153339 A1 US20050153339 A1 US 20050153339A1 US 485304 A US485304 A US 485304A US 2005153339 A1 US2005153339 A1 US 2005153339A1 Authority US United States Prior art keywords sequence vector sequences shine seq Prior art date Legal status (The legal status is an assumption and is not a legal conclusion. Google Patents US20050153339A1 - Modified shine-dalgarno sequences and methods of use thereof No commercial reproduction, distribution, display or performance rights in this work are provided.US20050153339A1 - Modified shine-dalgarno sequences and methods of use thereof The Shine-Dalgarno sequence of riboswitch-regulated single mRNAs shows ligand-dependent accessibility bursts. The authors declare no competing financial interests. All authors were involved in interpreting and discussing the results, and participated in writing the paper. wrote and implemented scripts for burst analysis. designed, conducted and analysed the in vitro translation experiments and SiM-KARTS experiments performed in the presence of blocking strand. wrote and implemented the scripts for burst analysis. designed, conducted and analysed the SiM-KARTS experiments. Wedekind) to N.G.W.Īuthor contributions: A.J.R. This research was funded by NIH grants GM062357, GM098023 and a sub-contract on GM063162 (PI: J. Hoff for performing the Fluorescence correlation spectroscopy measurements. We thank the Single Molecule Analysis in Real-Time (SMART) Center of the University of Michigan, seeded by NSF MRI-R2-ID award DBI-0959823 to N.G.W., as well as J.D. Suddala for careful reading of the manuscript, and G. Received 18 September 2015 Accepted 21 October 2015 Published 19 January 2016. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. This work is licensed under a Creative Commons Attribution 4.0 International License. Such complex ligand sensing by individual mRNA molecules rationalizes the nuanced ligand response observed during bulk mRNA translation. In addition, ligand-jump experiments reveal imperfect riboswitching of single mRNA molecules. Addition of preQ_1 decreases the lifetime of the SD’s high-accessibility (bursting) state and prolongs the time between bursts. Spike train analysis reveals that individual mRNA molecules alternate between two conformational states, distinguished by ‘bursts’ of probe binding associated with increased SD sequence accessibility. Here we develop Single Molecule Kinetic Analysis of RNA Transient Structure (SiM-KARTS) to investigate the ligand-dependent accessibility of the SD sequence of an mRNA hosting the 7-aminomethyl-7-deazaguanine (preQ_1)-sensing riboswitch. It is generally thought that this regulation originates from occlusion of the Shine-Dalgarno (SD) sequence upon ligand binding however, little direct evidence exists. In response to intracellular signals in Gram-negative bacteria, translational riboswitches-commonly embedded in messenger RNAs (mRNAs)-regulate gene expression through inhibition of translation initiation.
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