PMID- 22519888
OWN - NLM
STAT- MEDLINE
DCOM- 20121023
LR  - 20211021
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 84
IP  - 11
DP  - 2012 Jun 5
TI  - Engineered allosteric ribozymes that sense the bacterial second messenger cyclic 
      diguanosyl 5'-monophosphate.
PG  - 4935-41
LID - 10.1021/ac300415k [doi]
AB  - A series of allosteric ribozymes that respond to the bacterial second messenger 
      cyclic diguanosyl-5'-monophosphate (c-di-GMP) have been created by using in vitro 
      selection. An RNA library was generated by using random-sequence bridges to join 
      a hammerhead self-cleaving ribozyme to an aptamer from a natural c-di-GMP 
      riboswitch. Specific bridge sequences, called communication modules, emerged 
      through two in vitro selection efforts that either activate or inhibit ribozyme 
      self-cleavage upon ligand binding to the aptamer. Representative RNAs were found 
      that exhibit EC(50) (half-maximal effective concentration) values for c-di-GMP as 
      low as 90 nM and IC(50) (half-maximal inhibitory concentration) values as low as 
      180 nM. The allosteric RNAs display molecular recognition characteristics that 
      mimic the high discriminatory ability of the natural aptamer. Some engineered 
      RNAs operate with ribozyme rate constants approaching that of the parent 
      hammerhead ribozyme. By use of these allosteric ribozymes, cytoplasmic 
      concentrations of c-di-GMP in three mutant strains of Escherichia coli were 
      quantitatively estimated from cell lysates. Our findings demonstrate that 
      engineered c-di-GMP-sensing ribozymes can be used as convenient tools to monitor 
      c-di-GMP levels from complex biological or chemical samples. Moreover, these 
      ribozymes could be employed in high-throughput screens to identify compounds that 
      trigger c-di-GMP riboswitch function.
FAU - Gu, Hongzhou
AU  - Gu H
AD  - Department of Molecular, Cellular and Developmental Biology, Yale University, New 
      Haven, Connecticut, United States.
FAU - Furukawa, Kazuhiro
AU  - Furukawa K
FAU - Breaker, Ronald R
AU  - Breaker RR
LA  - eng
GR  - P01 GM022778/GM/NIGMS NIH HHS/United States
GR  - HHMI/Howard Hughes Medical Institute/United States
GR  - GM022778/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120521
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Aptamers, Nucleotide)
RN  - 0 (RNA, Catalytic)
RN  - 0 (Riboswitch)
RN  - 0 (hammerhead ribozyme)
RN  - 61093-23-0 (bis(3',5')-cyclic diguanylic acid)
RN  - 63231-63-0 (RNA)
RN  - H2D2X058MU (Cyclic GMP)
SB  - IM
MH  - Allosteric Regulation
MH  - Aptamers, Nucleotide/chemistry
MH  - Cyclic GMP/*analogs & derivatives/analysis/metabolism
MH  - Escherichia coli/genetics/*metabolism
MH  - Nucleic Acid Conformation
MH  - Protein Engineering
MH  - RNA/chemistry
MH  - RNA, Catalytic/*chemistry/genetics/metabolism
MH  - Riboswitch/physiology
MH  - Second Messenger Systems/*physiology
PMC - PMC4140410
MID - NIHMS379301
EDAT- 2012/04/24 06:00
MHDA- 2012/10/24 06:00
PMCR- 2014/08/21
CRDT- 2012/04/24 06:00
PHST- 2012/04/24 06:00 [entrez]
PHST- 2012/04/24 06:00 [pubmed]
PHST- 2012/10/24 06:00 [medline]
PHST- 2014/08/21 00:00 [pmc-release]
AID - 10.1021/ac300415k [doi]
PST - ppublish
SO  - Anal Chem. 2012 Jun 5;84(11):4935-41. doi: 10.1021/ac300415k. Epub 2012 May 21.