Triplex RNA motif binds cellular cGMP after expression in mammalian cells
The transmission of signals within cells is dependent on cyclic guanosine monophosphate (cGMP) as an important secondary messenger. German scientists have now developed an RNA that binds cGMP. As reported in the journal Angewandte Chemie, it is possible to suppress the cGMP signal cascade in genetically modified cells that produce this RNA.
cGMP plays an important role in processes such as the relaxation of the smooth muscle tissue in blood vessels and consequently in the regulation of blood pressure. Malfunction of the cGMP signaling pathway may be related to cardiovascular disease.
Experimental manipulation of the endogenous cGMP levels in cells should lead to a better understanding of the spatial and temporal dynamics involved, as well as the functionality of cGMP. While there are many ways to stimulate cGMP, including the use of nitrogen monoxide (NO), researchers have thus far not had a means to artificially lower cellular cGMP concentration.
Scientists from the Universities of Stuttgart and Tübingen have now developed a method by which they can “trap” cGMP molecules in cells. To achieve this they genetically modified the cells to produce specially designed RNA molecules that bind cGMP.
RNA, ribonucleic acid, is familiar to us as a building block of ribosomes, an amino acid transporter, and as messenger RNA, which copies blueprints from DNA and transports them to the ribosomes, where protein synthesis takes place. Further physiological roles have also now been found, such as catalytically active RNAs or RNAs that regulate gene expression by binding to complementary sequences. In addition, there are riboswitches, sequences in the messenger RNA that bind low-molecular metabolites and thus regulate gene expression.
A team led by Stuttgart chemist Clemens Richert and Tübingen biochemist Robert Feil has now successfully used specially developed RNA sequences to reduce the concentration of small molecules capable of base pairing in cells. To achieve this, the Stuttgart chemists developed a special folding motif that binds cGMP. The structure is based on a triple strand of RNA, known as a triplex. One of the three strands forms a loop that frames the binding cavity for cGMP. This motif is repeated multiple times in a long continuous sequence, so the researchers named their RNA construct “endless”.
In order to test the functionality of the “endless” construct in living cells, the biochemists in Tuebingen produced an artificial gene that codes for the “endless” RNA, and introduced it into cells obtained from the blood vessels of mice. This is a well-established model for the study of cGMP signaling pathways. In these cells, NO triggers signal cascades transmitted by cGMP. In cells that expressed “endless”, these cascades were suppressed and the cGMP level was significantly lower than in control cells. The “endless” RNA acts as a sink for cGMP and should be very useful in further research into the physiological role of cGMP.
About the Author
Clemens Richert is a synthetic organic chemist and Chair of Biological Chemistry at the University of Stuttgart. His research focuses on functional nucleic acids. He is also the chairman of the German Nucleic Acid Chemistry Society, DNG e.V. (http://dnarna.de).
Author: Clemens Richert, Universität Stuttgart (Germany), http://chip.chemie.uni-stuttgart.de/
Title: Endless: A Purine Binding Motif that Can Be Expressed in Cells
Angewandte Chemie International Edition, Permalink to the article: http://dx.doi.org/10.1002/anie.201403579
Clemens Richert | Angewandte Chemie
Locusts provide insight into brain response to stimuli, senses
28.04.2015 | Washington University in St. Louis
Discovery of an unexpected function of a protein linked to neurodegenerative diseases
28.04.2015 | Institute for Research in Biomedicine (IRB Barcelona)
KAIST researchers published an article on the development of a novel technique to precisely track the 3-D positions of optically-trapped particles having complicated geometry in high speed in the April 2015 issue of Optica.
Daejeon, Republic of Korea, April 23, 2015--Optical tweezers have been used as an invaluable tool for exerting micro-scale force on microscopic particles and...
A very small and rare species of shark is swimming its way through scientific literature. But don't worry, the chances of this inches-long vertebrate biting...
Ever since computers have been small enough to be fixtures on desks and laps, their central processing has functioned something like an atomic Etch A Sketch, with electromagnetic fields pushing data bits into place to encode data.
Unfortunately, the same drawbacks and perils of the mechanical sketch board have been just as pervasive in computing: making a change often requires starting...
How is lightning initiated in thunderclouds? This is difficult to answer - how do you measure electric fields inside large, dangerously charged clouds? It was discovered, more or less by coincidence, that cosmic rays provide suitable probes to measure electric fields within thunderclouds. This surprising finding is published in Physical Review Letters on April 24th. The measurements were performed with the LOFAR radio telescope located in the Netherlands.
How is lightning initiated in thunderclouds? This is difficult to answer - how do you measure electric fields inside large, dangerously charged clouds? It was...
Max Planck researcher Buhalqem Mamtimin determines how much nitrogen oxide is released into the atmosphere from agriculturally used oases.
In order to make statements about current and future air pollution, scientists use models which simulate the Earth’s atmosphere. A lot of information such as...
23.04.2015 | Event News
23.04.2015 | Event News
13.04.2015 | Event News
28.04.2015 | Press release
28.04.2015 | Power and Electrical Engineering
28.04.2015 | Earth Sciences