GlmS glucosamine-6-phosphate activated ribozyme

Last updated
glmS glucosamine-6-phosphate activated ribozyme
GlmS ribozyme secondary structure.jpg
Identifiers
SymbolglmS
Rfam RF00234
Other data
RNA type Cis-reg; riboswitch
Domain(s) Bacteria
SO SO:0000035
PDB structures PDBe
A 3D representation of the GlmS ribozyme. This is a view of the GlmS ribozyme bound to its catalytic cofactor. PDB 2nz4 EBI.jpg
A 3D representation of the GlmS ribozyme. This is a view of the GlmS ribozyme bound to its catalytic cofactor.
A 3D representation of the GlmS ribozyme. This view shows the pre-cleavage state of the Thermoanaerobacter tengcongensis glmS ribozyme bound to glucose-6-phosphate. PDB 2ho7 EBI.png
A 3D representation of the GlmS ribozyme. This view shows the pre-cleavage state of the Thermoanaerobacter tengcongensis glmS ribozyme bound to glucose-6-phosphate.

The glucosamine-6-phosphate riboswitch ribozyme (glmS ribozyme) is an RNA structure that resides in the 5' untranslated region (UTR) of the mRNA transcript of the glmS gene. This RNA regulates the glmS gene by responding to concentrations of a specific metabolite, glucosamine-6-phosphate (GlcN6P), in addition to catalyzing a self-cleaving chemical reaction upon activation. [3] This cleavage leads to the degradation of the mRNA that contains the ribozyme, and lowers production of GlcN6P. [4] The glmS gene encodes for an enzyme glutamine-fructose-6-phosphate amidotransferase, which catalyzes the formation of GlcN6P, a compound essential for cell wall biosynthesis, from fructose-6-phosphate and glutamine. [3] Thus, when GlcN6P levels are high, the glmS ribozyme is activated and the mRNA transcript is degraded but in the absence of GlcN6P the gene continues to be translated into glutamine-fructose-6-phosphate amidotransferase and GlcN6P is produced. GlcN6P is a cofactor for this cleavage reaction, as it directly participates as an acid-base catalyst. [5] This RNA is the first riboswitch also found to be a self-cleaving ribozyme and, like many others, was discovered using a bioinformatics approach. [6]

Contents

Structure

The structure of the glmS ribozyme was first determined by X-ray crystallography in 2006. [1] [2] The tertiary structure of this RNA is characterized by three coaxial stacked helices, packed side by side. [2] The ribozyme core contains a double pseudoknotted structure, which places the central helix P2.1 such that the scissile phosphate is nestled by the major groove. [2] The major groove of the adjacent helix P2.2 is involved in metabolite binding and the scissile phosphate is attached to the 5' end of the helix. [2] The roof of the active site is characterized by conserved base triples, which connect P2.1 and P2.2 stacks and the floor consists of a non-conserved G-U pair, which are splayed apart. [2] By examining superimposition of ribozyme structures in a pre-cleavage state, metabolite bound state and post cleavage state, it was determined there is no gross conformational change upon metabolite binding, which is indicative of a preorganized active site that depends on GlcN6P as a cofactor, not an allosteric activator. [2] The cofactor is bound in a solvent-accessible pocket and the structure suggests that the amine group of GlcN6P is involved in the catalytic process. [2] [1] [7] [8]

Cleavage Requirements

Although glmS ribozyme performs self-cleavage upon binding GlcN6P, the glmS ribozyme active site does not undergo conformational change upon binding GlcN6P. [2] Instead, the active site is pre-formed and glmS ribozyme activity is triggered by the introduction of a functional group on GlcN6P that is necessary for catalysis. [2] [8] As mentioned above, the amine group on GlcN6P is thought to be the catalytically involved functional group, which is supported by studies that evaluated glmS ribozyme activity using different ligands, under physiologic conditions. [9] For example, these experiments found that incubation with Glc6P (no amine group) inhibits glmS self-cleavage while incubation with GlcN (available amine group) stimulates cleavage, though not to the extent that GlcN6P does. [9] These and other findings indicate that the glmS ribozyme is a unique class of ribozyme, and that its discovery is further evidence of the catalytic capacity of RNA. [8]

See also

Related Research Articles

Peptidoglycan or murein is a polymer consisting of sugars and amino acids that forms a mesh-like peptidoglycan layer outside the plasma membrane of most bacteria, forming the cell wall. The sugar component consists of alternating residues of β-(1,4) linked N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM). Attached to the N-acetylmuramic acid is a peptide chain of three to five amino acids. The peptide chain can be cross-linked to the peptide chain of another strand forming the 3D mesh-like layer. Peptidoglycan serves a structural role in the bacterial cell wall, giving structural strength, as well as counteracting the osmotic pressure of the cytoplasm. Peptidoglycan is also involved in binary fission during bacterial cell reproduction.

Ribozyme

Ribozymes are RNA molecules that have the ability to catalyze specific biochemical reactions, including RNA splicing in gene expression, similar to the action of protein enzymes. The 1982 discovery of ribozymes demonstrated that RNA can be both genetic material and a biological catalyst, and contributed to the RNA world hypothesis, which suggests that RNA may have been important in the evolution of prebiotic self-replicating systems. The most common activities of natural or in vitro-evolved ribozymes are the cleavage or ligation of RNA and DNA and peptide bond formation. Within the ribosome, ribozymes function as part of the large subunit ribosomal RNA to link amino acids during protein synthesis. They also participate in a variety of RNA processing reactions, including RNA splicing, viral replication, and transfer RNA biosynthesis. Examples of ribozymes include the hammerhead ribozyme, the VS ribozyme, Leadzyme and the hairpin ribozyme.

Riboswitch

In molecular biology, a riboswitch is a regulatory segment of a messenger RNA molecule that binds a small molecule, resulting in a change in production of the proteins encoded by the mRNA. Thus, an mRNA that contains a riboswitch is directly involved in regulating its own activity, in response to the concentrations of its effector molecule. The discovery that modern organisms use RNA to bind small molecules, and discriminate against closely related analogs, expanded the known natural capabilities of RNA beyond its ability to code for proteins, catalyze reactions, or to bind other RNA or protein macromolecules.

Glutamine synthetase

Glutamine synthetase (GS) is an enzyme that plays an essential role in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine:

Hammerhead ribozyme

The hammerhead ribozyme is an RNA motif that catalyzes reversible cleavage and ligation reactions at a specific site within an RNA molecule. It is one of several catalytic RNAs (ribozymes) known to occur in nature. It serves as a model system for research on the structure and properties of RNA, and is used for targeted RNA cleavage experiments, some with proposed therapeutic applications. Named for the resemblance of early secondary structure diagrams to a hammerhead shark, hammerhead ribozymes were originally discovered in two classes of plant virus-like RNAs: satellite RNAs and viroids. They have subsequently been found to be widely dispersed within many forms of life.

Hairpin ribozyme

The hairpin ribozyme is a small section of RNA that can act as a ribozyme. Like the hammerhead ribozyme it is found in RNA satellites of plant viruses. It was first identified in the minus strand of the tobacco ringspot virus (TRSV) satellite RNA where it catalyzes self-cleavage and joining (ligation) reactions to process the products of rolling circle virus replication into linear and circular satellite RNA molecules. The hairpin ribozyme is similar to the hammerhead ribozyme in that it does not require a metal ion for the reaction.

Cobalamin riboswitch

Cobalamin riboswitch is a cis-regulatory element which is widely distributed in 5' untranslated regions of vitamin B12 (Cobalamin) related genes in bacteria.

Amidophosphoribosyltransferase

Amidophosphoribosyltransferase (ATase), also known as glutamine phosphoribosylpyrophosphate amidotransferase (GPAT), is an enzyme responsible for catalyzing the conversion of 5-phosphoribosyl-1-pyrophosphate (PRPP) into 5-phosphoribosyl-1-amine (PRA), using the amine group from a glutamine side-chain. This is the committing step in de novo purine synthesis. In humans it is encoded by the PPAT gene. ATase is a member of the purine/pyrimidine phosphoribosyltransferase family.

Leadzyme

Leadzyme is a small ribozyme (catalytic RNA), which catalyzes the cleavage of a specific phosphodiester bond. It was discovered using an in-vitro evolution study where the researchers were selecting for RNAs that specifically cleaved themselves in the presence of lead. However, since then, it has been discovered in several natural systems. Leadzyme was found to be efficient and dynamic in the presence of micromolar concentrations of lead ions. Unlike in other small self-cleaving ribozymes, other divalent metal ions cannot replace Pb2+ in the leadzyme. Due to obligatory requirement for a lead, the ribozyme is called a metalloribozyme.

VS ribozyme

The Varkud satellite (VS) ribozyme is an RNA enzyme that carries out the cleavage of a phosphodiester bond.

Hepatitis delta virus ribozyme

The hepatitis delta virus (HDV) ribozyme is a non-coding RNA found in the hepatitis delta virus that is necessary for viral replication and is the only known human virus that utilizes ribozyme activity to infect its host. The ribozyme acts to process the RNA transcripts to unit lengths in a self-cleavage reaction during replication of the hepatitis delta virus, which is thought to propagate by a double rolling circle mechanism. The ribozyme is active in vivo in the absence of any protein factors and was the fastest known naturally occurring self-cleaving RNA at the time of its discovery.

GlmZ RNA Small non-coding RNA (ncRNA)

GlmZ is a small non-coding RNA (ncRNA). It is the functional product of a gene which is not translated into protein.

Uridine diphosphate <i>N</i>-acetylglucosamine Chemical compound

Uridine diphosphate N-acetylglucosamine or UDP-GlcNAc is a nucleotide sugar and a coenzyme in metabolism. It is used by glycosyltransferases to transfer N-acetylglucosamine residues to substrates. D-Glucosamine is made naturally in the form of glucosamine-6-phosphate, and is the biochemical precursor of all nitrogen-containing sugars. To be specific, glucosamine-6-phosphate is synthesized from fructose 6-phosphate and glutamine as the first step of the hexosamine biosynthesis pathway. The end-product of this pathway is UDP-GlcNAc, which is then used for making glycosaminoglycans, proteoglycans, and glycolipids.

N-acetylglucosamine-6-phosphate deacetylase

In enzymology, N-acetylglucosamine-6-phosphate deacetylase (EC 3.5.1.25), also known as GlcNAc-6-phosphate deacetylase or NagA, is an enzyme that catalyzes the deacetylation of N-acetylglucosamine-6-phosphate (GlcNAc-6-P) to glucosamine-6-phosphate (GlcN-6-P):

Glucosamine-phosphate N-acetyltransferase

In enzymology, glucosamine-phosphate N-acetyltransferase (GNA) is an enzyme that catalyzes the transfer of an acetyl group from acetyl-CoA to the primary amine in glucosamide-6-phosphate, generating a free CoA and N-acetyl-D-glucosamine-6-phosphate.

GFPT1

Glucosamine—fructose-6-phosphate aminotransferase isomerizing 1 is an enzyme that in humans is encoded by the GFPT1 gene.

Fluoride riboswitch Fluoride-binding RNA structure

The fluoride riboswitch is a conserved RNA structure identified by bioinformatics in a wide variety of bacteria and archaea. These RNAs were later shown to function as riboswitches that sense fluoride ions. These "fluoride riboswitches" increase expression of downstream genes when fluoride levels are elevated, and the genes are proposed to help mitigate the toxic effects of very high levels of fluoride.

Scott A. Strobel is the Henry Ford II professor of molecular biophysics and biochemistry and a professor of chemistry at Yale University. He is the vice provost for Science Initiatives and vice president for West Campus Planning & Program Development. An educator and researcher, he has led a number of Yale initiatives over the past two decades, Strobel was named in 2019 as Yale's next provost.

Twister ribozyme

The twister ribozyme is a catalytic RNA structure capable of self-cleavage. The nucleolytic activity of this ribozyme has been demonstrated both in vivo and in vitro and has one of the fastest catalytic rates of naturally occurring ribozymes with similar function. The twister ribozyme is considered to be a member of the small self-cleaving ribozyme family which includes the hammerhead, hairpin, hepatitis delta virus (HDV), Varkud satellite (VS), and glmS ribozymes.

Hatchet ribozyme

Background: The hatchet ribozyme is an RNA structure that catalyzes its own cleavage at a specific site. In other words, it is a self-cleaving ribozyme. Hatchet ribozymes were discovered by a bioinformatics strategy as RNAs Associated with Genes Associated with Twister and Hammerhead ribozymes, or RAGATH.

References

  1. 1 2 3 Cochrane JC, Lipchock SV, Strobel SA (2007). "Structural investigation of the GlmS ribozyme bound to its catalytic cofactor". Chem. Biol. 14 (1): 97–105. doi:10.1016/j.chembiol.2006.12.005. PMC   1847778 . PMID   17196404.
  2. 1 2 3 4 5 6 7 8 9 10 Klein DJ, Ferré-D'Amaré AR (2006). "Structural basis of glmS ribozyme activation by glucosamine-6-phosphate". Science. 313 (5794): 1752–1756. doi:10.1126/science.1129666. PMID   16990543. S2CID   37470517.
  3. 1 2 Winkler, WC; Nahvi A; Roth A; Collins JA; Breaker RR (2004). "Control of gene expression by a natural metabolite-responsive ribozyme". Nature. 428 (6980): 281–286. doi:10.1038/nature02362. PMID   15029187. S2CID   4301164.
  4. Collins, JA; Irnov I; Baker S; Winkler WC (2007). "Mechanism of mRNA destabilization by the glmS ribozyme". Genes Dev. 21 (24): 3356–3368. doi:10.1101/gad.1605307. PMC   2113035 . PMID   18079181.
  5. Viladoms, J. L.; Fedor, M. J. (2012). "TheglmSRibozyme Cofactor is a General Acid–Base Catalyst". Journal of the American Chemical Society. 134 (46): 19043–19049. doi:10.1021/ja307021f. PMC   3504194 . PMID   23113700.
  6. Barrick JE, Corbino KA, Winkler WC, et al. (April 2004). "New RNA motifs suggest an expanded scope for riboswitches in bacterial genetic control". Proc. Natl. Acad. Sci. U.S.A. 101 (17): 6421–6426. doi: 10.1073/pnas.0308014101 . PMC   404060 . PMID   15096624.
  7. Jansen JA, McCarthy TJ, Soukup GA, Soukup JK (2006). "Backbone and nucleobase contacts to glucosamine-6-phosphate in the glmS ribozyme". Nat Struct Mol Biol. 13 (6): 517–523. doi:10.1038/nsmb1094. PMID   16699515. S2CID   7263581.
  8. 1 2 3 Hampel KJ, Tinsley MM (2006). "Evidence for preorganization of the glmS ribozyme ligand binding pocket". Biochemistry. 45 (25): 7861–7871. doi:10.1021/bi060337z. PMID   16784238.
  9. 1 2 McCarthy TJ, Floy SA, Jansen JA, Soukup JK, Soukup GA (2006). "Ligand requirements for glmS ribozyme self-cleavage". Chemistry & Biology. 12 (11): 1221–1226. doi: 10.1016/j.chembiol.2005.09.006 . PMID   16298301.