Messenger RNA decapping

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Schematic representation of deadenylation-independent decapping in S. cerevisiae Dekeping, nezalezhnii vid deadeniliuvannia, u drizhdzhiv.png
Schematic representation of deadenylation-independent decapping in S. cerevisiae

The process of messenger RNA decapping consists of hydrolysis of the 5' cap structure on the RNA exposing a 5' monophosphate. In eukaryotes, this 5' monophosphate is a substrate for the 5' exonuclease Xrn1 [1] and the mRNA is quickly destroyed. There are many situations which may lead to the removal of the cap, some of which are discussed below. [2]

Contents

In prokaryotes, the initial mRNA transcript naturally possesses a 5'-triphosphate group after bacterial transcription; the enzyme RppH removes a pyrophosphate molecule from the 5' end, converting the 5'-triphosphate to a 5'-monophosphate, triggering mRNA degradation by ribonucleases. [3] [4]

Translation and decay

Inside cells, there is a balance between the processes of translation and mRNA decay. [2] Messages which are being actively translated are bound by polysomes and the eukaryotic initiation factors eIF-4E and eIF-4G (in eukaryotes). This blocks access to the cap by the decapping enzyme DCP2 and protects the mRNA molecule. In nutrient-starvation conditions or viral infection, translation may be compromised and decapping is stimulated. This balance is reflected in the size and abundance of the cytoplasmic structures known as P-bodies. [5] [6]

Specific decay pathways

A number of specific decay pathways exist that recognize aberrant messages and promote their decapping. Nonsense mediated decay recognizes premature stop codons and promotes decapping as well as decay by the exosome. Certain classes of miRNA have also been shown to stimulate decapping.

Related Research Articles

Adenosine triphosphate Chemical compound

Adenosine triphosphate (ATP) is an organic compound and hydrotrope that provides energy to drive many processes in living cells, such as muscle contraction, nerve impulse propagation, condensate dissolution, and chemical synthesis. Found in all known forms of life, ATP is often referred to as the "molecular unit of currency" of intracellular energy transfer. When consumed in metabolic processes, it converts either to adenosine diphosphate (ADP) or to adenosine monophosphate (AMP). Other processes regenerate ATP so that the human body recycles its own body weight equivalent in ATP each day. It is also a precursor to DNA and RNA, and is used as a coenzyme.

Messenger RNA RNA that is read by the ribosome to produce a protein

In molecular biology, messenger ribonucleic acid (mRNA) is a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene, and is read by a ribosome in the process of synthesizing a protein.

Nucleotide Biological molecules that form the building blocks of nucleic acids

Nucleotides are organic molecules consisting of a nucleoside and a phosphate. They serve as monomeric units of the nucleic acid polymers – deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), both of which are essential biomolecules within all life-forms on Earth. Nucleotides are obtained in the diet and are also synthesized from common nutrients by the liver.

Pyrophosphate Chemical compound

In chemistry, pyrophosphates are phosphorus oxyanions that contain two phosphorus atoms in a P-O-P linkage. A number of pyrophosphate salts exist, such as Na2H2P2O7. Often pyrophosphates are called diphosphates. The parent pyrophosphates are derived from partial or complete neutralization of pyrophosphoric acid. Important salts include disodium pyrophosphate and tetrasodium pyrophosphate. The pyrophosphate bond is also sometimes referred to as a phosphoanhydride bond, a naming convention which emphasizes the loss of water that occurs when two phosphates form a new P-O-P bond, and which mirrors the nomenclature for anhydrides of carboxylic acids. Pyrophosphates are found in ATP and other nucleotide triphosphates, which are very important in biochemistry.

Guanosine triphosphate Chemical compound

Guanosine-5'-triphosphate (GTP) is a purine nucleoside triphosphate. It is one of the building blocks needed for the synthesis of RNA during the transcription process. Its structure is similar to that of the guanosine nucleoside, the only difference being that nucleotides like GTP have phosphates on their ribose sugar. GTP has the guanine nucleobase attached to the 1' carbon of the ribose and it has the triphosphate moiety attached to ribose's 5' carbon.

Polyadenylation is the addition of a poly(A) tail to an RNA transcript, typically a messenger RNA (mRNA). The poly(A) tail consists of multiple adenosine monophosphates; in other words, it is a stretch of RNA that has only adenine bases. In eukaryotes, polyadenylation is part of the process that produces mature mRNA for translation. In many bacteria, the poly(A) tail promotes degradation of the mRNA. It, therefore, forms part of the larger process of gene expression.

In molecular biology, the five-prime cap is a specially altered nucleotide on the 5′ end of some primary transcripts such as precursor messenger RNA. This process, known as mRNA capping, is highly regulated and vital in the creation of stable and mature messenger RNA able to undergo translation during protein synthesis. Mitochondrial mRNA and chloroplastic mRNA are not capped.

Exonuclease

Exonucleases are enzymes that work by cleaving nucleotides one at a time from the end (exo) of a polynucleotide chain. A hydrolyzing reaction that breaks phosphodiester bonds at either the 3′ or the 5′ end occurs. Its close relative is the endonuclease, which cleaves phosphodiester bonds in the middle (endo) of a polynucleotide chain. Eukaryotes and prokaryotes have three types of exonucleases involved in the normal turnover of mRNA: 5′ to 3′ exonuclease (Xrn1), which is a dependent decapping protein; 3′ to 5′ exonuclease, an independent protein; and poly(A)-specific 3′ to 5′ exonuclease.

Primary transcript RNA produced by transcription

A primary transcript is the single-stranded ribonucleic acid (RNA) product synthesized by transcription of DNA, and processed to yield various mature RNA products such as mRNAs, tRNAs, and rRNAs. The primary transcripts designated to be mRNAs are modified in preparation for translation. For example, a precursor mRNA (pre-mRNA) is a type of primary transcript that becomes a messenger RNA (mRNA) after processing.

P-bodies, or processing bodies are distinct foci formed by phase separation within the cytoplasm of the eukaryotic cell consisting of many enzymes involved in mRNA turnover. P-bodies are highly conserved structures and have been observed in somatic cells originating from vertebrates and invertebrates, plants and yeast. To date, P-bodies have been demonstrated to play fundamental roles in general mRNA decay, nonsense-mediated mRNA decay, adenylate-uridylate-rich element mediated mRNA decay, and microRNA (miRNA) induced mRNA silencing. Not all mRNAs which enter P-bodies are degraded, as it has been demonstrated that some mRNAs can exit P-bodies and re-initiate translation. Purification and sequencing of the mRNA from purified processing bodies showed that these mRNAs are largely translationally repressed upstream of translation initiation and are protected from 5' mRNA decay.

The 5' cap of eukaryotic messenger RNA is bound at all times by various cap-binding complexes (CBCs).

XRN1 (gene)

5′-3′ exoribonuclease 1 (Xrn1) is a protein that in humans is encoded by the XRN1 gene. Xrn1 hydrolyses RNA in the 5′ to 3′ direction.

The degradosome is a multiprotein complex present in most bacteria that is involved in the processing of ribosomal RNA and the degradation of messenger RNA and is regulated by Non-coding RNA. It contains the proteins RNA helicase B, RNase E and Polynucleotide phosphorylase.

In enzymology, a polynucleotide 5'-phosphatase (RNA 5'-triphosphatase, RTPase, EC 3.1.3.33) is an enzyme that catalyzes the chemical reaction

DCP2

mRNA-decapping enzyme 2 is a protein that in humans is encoded by the DCP2 gene.

Decapping complex Eukaryotic protein complex that removes the 5 cap on mRNA

The mRNA decapping complex is a protein complex in eukaryotic cells responsible for removal of the 5' cap. The active enzyme of the decapping complex is the bilobed Nudix family enzyme Dcp2, which hydrolyzes 5' cap and releases 7mGDP and a 5'-monophosphorylated mRNA. This decapped mRNA is inhibited for translation and will be degraded by exonucleases. The core decapping complex is conserved in eukaryotes. Dcp2 is activated by Decapping Protein 1 (Dcp1) and in higher eukaryotes joined by the scaffold protein VCS. Together with many other accessory proteins, the decapping complex assembles in P-bodies in the cytoplasm.

Nudix hydrolase

Nudix hydrolases are a superfamily of hydrolytic enzymes capable of cleaving nucleoside diphosphates linked to x, hence their name. The reaction yields nucleoside monophosphate (NMP) plus X-P. Substrates hydrolysed by nudix enzymes comprise a wide range of organic pyrophosphates, including nucleoside di- and triphosphates, dinucleoside and diphosphoinositol polyphosphates, nucleotide sugars and RNA caps, with varying degrees of substrate specificity. Enzymes of the Nudix superfamily are found in all types of organisms, including eukaryotes, bacteria and archaea.

Cryptic unstable transcripts (CUTs) are a subset of non-coding RNAs (ncRNAs) that are produced from intergenic and intragenic regions. CUTs were first observed in S. cerevisiae yeast models and are found in most eukaryotes. Some basic characteristics of CUTs include a length of around 200–800 base pairs, a 5' cap, poly-adenylated tail, and rapid degradation due to the combined activity of poly-adenylating polymerases and exosome complexes. CUT transcription occurs through RNA Polymerase II and initiates from nucleosome-depleted regions, often in an antisense orientation. To date, CUTs have a relatively uncharacterized function but have been implicated in a number of putative gene regulation and silencing pathways. Thousands of loci leading to the generation of CUTs have been described in the yeast genome. Additionally, stable uncharacterized transcripts, or SUTs, have also been detected in cells and bear many similarities to CUTs but are not degraded through the same pathways.

Roy R. Parker is a biochemist who has been an active investigator in science since the 1970s. He is currently a Distinguished Professor of Chemistry and Biochemistry and Cech-Leinwand Endowed Chair of Biochemistry at the University of Colorado Boulder. Throughout his life, Parker has contributed a vast degree of knowledge to research and studies of biochemistry. His current focus includes the biogenesis, function, and degradation of multiple forms of RNA in eukaryotes. Parker aims to use his research to understand how various diseases and pathologies result from abnormalities in RNA. In 2012, Parker was elected to the National Academy of Sciences in Biochemistry.

In molecular biology, the NAD+ five-prime cap refers to a molecule of nicotinamide adenine dinucleotide (NAD+), a nucleoside-containing metabolite, covalently bonded the 5’ end of cellular mRNA. While the more common methylated guanosine (m7G) cap is added to RNA by a capping complex that associates with RNA polymerase II, the NAD cap is added during transcriptional initiation by the RNA polymerase itself, acting as a non-canonical initiating nucleotide (NCIN). As such, while m7G capping can only occur in organisms possessing specialized capping complexes, because NAD capping is performed by RNAP itself, it is hypothesized to occur in most, if not all, organisms.

References

  1. Poole, Toni L.; Stevens, Audrey (June 1997). "Structural Modifications of RNA Influence the 5′ Exoribonucleolytic Hydrolysis by XRN1 and HKE1 ofSaccharomyces cerevisiae" (PDF). Biochemical and Biophysical Research Communications. 235 (3): 799–805. doi:10.1006/bbrc.1997.6877. PMID   9207242.
  2. 1 2 Meyer, Sylke; Temme, Claudia; Wahle, Elmar (January 2004). "Messenger RNA Turnover in Eukaryotes: Pathways and Enzymes". Critical Reviews in Biochemistry and Molecular Biology. 39 (4): 197–216. doi:10.1080/10409230490513991. PMID   15596551.
  3. Deana, Atilio; Celesnik, Helena; Belasco, Joel G. (17 January 2008). "The bacterial enzyme RppH triggers messenger RNA degradation by 5′ pyrophosphate removal". Nature. 451 (7176): 355–358. doi:10.1038/nature06475. PMID   18202662.
  4. Hsieh, Ping-kun; Richards, Jamie; Liu, Quansheng; Belasco, Joel G. (22 April 2013). "Specificity of RppH-dependent RNA degradation in Bacillus subtilis". Proceedings of the National Academy of Sciences. 110 (22): 8864–8869. doi:10.1073/pnas.1222670110. PMC   3670361 . PMID   23610425.
  5. Sheth, Ujwal; Parker, Roy (2 May 2003). "Decapping and Decay of Messenger RNA Occur in Cytoplasmic Processing Bodies". Science. 300 (5620): 805–808. doi:10.1126/science.1082320. PMC   1876714 . PMID   12730603.
  6. Parker, Roy; Sheth, Ujwal (March 2007). "P Bodies and the Control of mRNA Translation and Degradation". Molecular Cell. 25 (5): 635–646. doi: 10.1016/j.molcel.2007.02.011 . PMID   17349952.