RNA editing is a molecular process through which some cells can make discrete changes to specific nucleotide sequences within an RNA molecule after it has been generated by RNA polymerase. RNA editing is relatively rare, and common forms of RNA processing (e.g. splicing, 5'-capping, and 3'-polyadenylation) are not usually included as editing. The editing events may include the insertion, deletion, and base substitution of nucleotides within the edited RNA molecule.
RNA editing has been observed in some tRNA, rRNA, mRNA, or miRNA molecules of eukaryotes and their viruses, archaea, and prokaryotes. RNA editing occurs in the cell nucleus and cytosol, as well as within mitochondria and plastids. In vertebrates, editing is rare and usually consists of a small number of changes to the sequence of affected molecules. In other organisms, extensive editing (pan-editing) can occur; in some cases the majority of nucleotides in a mRNA sequence may result from editing.
RNA-editing processes show great molecular diversity, and some appear to be evolutionarily recent acquisitions that arose independently. The diversity of RNA editing phenomena includes nucleobase modifications such as cytidine (C) to uridine (U) and adenosine (A) to inosine (I) deaminations, as well as non-templated nucleotide additions and insertions. RNA editing in mRNAs effectively alters the amino acid sequence of the encoded protein so that it differs from that predicted by the genomic DNA sequence.
Editing by insertion or deletionEdit
RNA editing through the addition and deletion of uracil has been found in kinetoplasts from the mitochondria of Trypanosoma brucei Because this may involve a large fraction of the sites in a gene, it is sometimes called "pan-editing" to distinguish it from topical editing of one or a few sites.
Pan-editing starts with the base-pairing of the unedited primary transcript with a guide RNA (gRNA), which contains complementary sequences to the regions around the insertion/deletion points. The newly formed double-stranded region is then enveloped by an editosome, a large multi-protein complex that catalyzes the editing. The editosome opens the transcript at the first mismatched nucleotide and starts inserting uridines. The inserted uridines will base-pair with the guide RNA, and insertion will continue as long as A or G is present in the guide RNA and will stop when a C or U is encountered. The inserted nucleotides cause a frameshift, and result in a translated protein that differs from its gene.
The mechanism of the editosome involves an endonucleolytic cut at the mismatch point between the guide RNA and the unedited transcript. The next step is catalyzed by one of the enzymes in the complex, a terminal U-transferase, which adds Us from UTP at the 3' end of the mRNA. The opened ends are held in place by other proteins in the complex. Another enzyme, a U-specific exoribonuclease, removes the unpaired Us. After editing has made mRNA complementary to gRNA, an RNA ligase rejoins the ends of the edited mRNA transcript. As a consequence, the editosome can edit only in a 3' to 5' direction along the primary RNA transcript. The complex can act on only a single guide RNA at a time. Therefore, a RNA transcript requiring extensive editing will need more than one guide RNA and editosome complex.
Editing by deaminationEdit
The editing involves cytidine deaminase that deaminates a cytidine base into a uridine base. An example of C-to-U editing is with the apolipoprotein B gene in humans. Apo B100 is expressed in the liver and apo B48 is expressed in the intestines. In the intestines, the mRNA has a CAA sequence edited to be UAA, a stop codon, thus producing the shorter B48 form. C-to-U editing often occurs in the mitochondrial RNA of flowering plants. Different plants have different degrees of C-to-U editing; for example, eight (8) editing events occur in mitochondria of the moss Funaria hygrometrica, whereas over 1,700 editing events occur in the lycophytes Isoetes engelmanii. C-to-U editing is performed by members of the pentatricopeptide repeat (PPR) protein family. Angiosperms have large PPR families, acting as trans -factors for cis -elements lacking a consensus sequence; Arabidopsis has around 450 members in its PPR family. There have been a number of discoveries of PPR proteins in both plastids and mitochondria.
Adenosine-to-inosine (A-to-I) modifications contribute to nearly 90% of all editing events in RNA. The deamination of adenosine is catalyzed by the double-stranded RNA-specific adenosine deaminase (ADAR), which typically acts on pre-mRNAs. The deamination of adenosine to inosine disrupts and destabilizes the dsRNA base pairing, therefore rendering that particular dsRNA less able to produce siRNA, which interferes with the RNAi pathway.
The wobble base pairing causes deaminated RNA to have a unique but different structure, which may be related to the inhibition of the initiation step of RNA translation. Studies have shown that I-RNA (RNA with many repeats of the I-U base pair) recruits methylases that are involved in the formation of heterochromatin and that this chemical modification heavily interferes with miRNA target sites. There is active research into the importance of A-to-I modifications and their purpose in the novel concept of epitranscriptomics, in which modifications are made to RNA that alter their function.
The development of high-throughput sequencing in recent years has allowed for the development of extensive databases for different modifications and edits of RNA. RADAR (Rigorously Annotated Database of A-to-I RNA editing) was developed in 2013 to catalog the vast variety of A-to-I sites and tissue-specific levels present in humans, mice, and flies. The addition of novel sites and overall edits to the database are ongoing. The level of editing for specific editing sites, e.g. in the filamin A transcript, is tissue-specific. The efficiency of mRNA-splicing is a major factor controlling the level of A-to-I RNA editing.
Alternative mRNA editingEdit
Alternative U-to-C mRNA editing was first reported in WT1 (Wilms Tumor-1) transcripts, and non-classic G-A mRNA changes were first observed in HNRNPK (heterogeneous nuclear ribonucleoprotein K) transcripts in both malignant and normal colorectal samples. The latter changes were also later seen alongside non-classic U-to-C alterations in brain cell TPH2 (tryptophan hydroxylase 2) transcripts. Although the reverse amination might be the simplest explanation for U-to-C changes, transamination and transglycosylation mechanisms have been proposed for plant U-to-C editing events in mitochondrial transcripts. A recent study reported novel G-to-A mRNA changes in WT1 transcripts at two hotspots, proposing the APOBEC3A (apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3A) as the enzyme implicated in this class of alternative mRNA editing. It was also shown that alternative mRNA changes were associated with canonical WT1 splicing variants, indicating their functional significance.
RNA editing in plant mitochondria and plastidsEdit
It has been shown in previous studies that the only types of RNA editing seen in the plants' mitochondria and plastids are conversion of C-to-U and U-to-C (very rare). RNA-editing sites are found mainly in the coding regions of mRNA, introns, and other non-translated regions. In fact, RNA editing can restore the functionality of tRNA molecules. The editing sites are found primarily upstream of mitochondrial or plastid RNAs. While the specific positions for C to U RNA editing events have been fairly well studied in both the mitochondrion and plastid, the identity and organization of all proteins comprising the editosome have yet to be established. Members of the expansive PPR protein family have been shown to function as trans-acting factors for RNA sequence recognition. Specific members of the MORF (Multiple Organellar RNA editing Factor) family are also required for proper editing at several sites. As some of these MORF proteins have been shown to interact with members of the PPR family, it is possible MORF proteins are components of the editosome complex. An enzyme responsible for the trans- or deamination of the RNA transcript remains elusive, though it has been proposed that the PPR proteins may serve this function as well.
RNA editing is essential for the normal functioning of the plant's translation and respiration activity. Editing can restore the essential base-pairing sequences of tRNAs, restoring functionality. It has also been linked to the production of RNA-edited proteins that are incorporated into the polypeptide complexes of the respiration pathway. Therefore, it is highly probable that polypeptides synthesized from unedited RNAs would not function properly and hinder the activity of both mitochondria and plastids.
C-to-U RNA editing can create start and stop codons, but it cannot destroy existing start and stop codons. A cryptic start codon is created when the codon ACG is edited to be AUG.
RNA editing in virusesEdit
RNA editing in viruses (i.e., measles, mumps, or parainfluenza) is used for stability and generation of protein variants. Viral RNAs are transcribed by a virus-encoded RNA-dependent RNA polymerase, which is prone to pausing and "stuttering" at certain nucleotide combinations. In addition, up to several hundred non-templated A's are added by the polymerase at the 3' end of nascent mRNA. These As help stabilize the mRNA. Furthermore, the pausing and stuttering of the RNA polymerase allows the incorporation of one or two Gs or As upstream of the translational codon. The addition of the non-templated nucleotides shifts the reading frame, which generates a different protein.
Origin and evolution of RNA editingEdit
The RNA-editing system seen in the animal may have evolved from mononucleotide deaminases, which have led to larger gene families that include the apobec-1 and adar genes. These genes share close identity with the bacterial deaminases involved in nucleotide metabolism. The adenosine deaminase of E. coli cannot deaminate a nucleoside in the RNA; the enzyme's reaction pocket is too small to for the RNA strand to bind to. However, this active site is widened by amino acid changes in the corresponding human analog genes, APOBEC1 and ADAR, allowing deamination. The gRNA-mediated pan-editing in trypanosome mitochondria, involving templated insertion of U residues, is an entirely different biochemical reaction. The enzymes involved have been shown in other studies to be recruited and adapted from different sources. But the specificity of nucleotide insertion via the interaction between the gRNA and mRNA is similar to the tRNA editing processes in the animal and Acanthamoeba mithochondria. Eukaryotic ribose methylation of rRNAs by guide RNA molecules is a similar form of modification.
Thus, RNA editing evolved more than once. Several adaptive rationales for editing have been suggested. Editing is often described as a mechanism of correction or repair to compensate for defects in gene sequences. However, in the case of gRNA-mediated editing, this explanation does not seem possible because if a defect happens first, there is no way to generate an error-free gRNA-encoding region, which presumably arises by duplication of the original gene region. This thinking leads to an evolutionary proposal called "constructive neutral evolution" in which the order of steps is reversed, with the gratuitous capacity for editing preceding the "defect". 31
RNA editing may be involved in RNA degradationEdit
A study looked at the involvement of RNA editing in RNA degradation. The researchers specifically looked at the interaction between ADAR and UPF1, an enzyme involved in the nonsense-mediated mRNA decay pathway (NMD). They found that ADAR and UPF1 are found within the suprasliceosome and they form a complex that leads to the down-regulation of specific genes. The exact mechanism or the exact pathways that these two are involved in are unknown at this time. The only fact that this research has shown is that they form a complex and down-regulate specific genes.
- Su AA, Randau L (August 2011). "A-to-I and C-to-U editing within transfer RNAs". Biochemistry. Biokhimiia. 76 (8): 932–7. doi:10.1134/S0006297911080098. PMID 22022967.
- Brennicke A, Marchfelder A, Binder S (June 1999). "RNA editing". FEMS Microbiology Reviews. 23 (3): 297–316. doi:10.1111/j.1574-6976.1999.tb00401.x. PMID 10371035.
- Benne R (April 1994). "RNA editing in trypanosomes". European Journal of Biochemistry. 221 (1): 9–23. doi:10.1111/j.1432-1033.1994.tb18710.x. PMID 7513284.
- Arts GJ, Benne R (June 1996). "Mechanism and evolution of RNA editing in kinetoplastida". Biochimica et Biophysica Acta. 1307 (1): 39–54. doi:10.1016/0167-4781(96)00021-8. PMID 8652667.
- Alfonzo JD, Thiemann O, Simpson L (October 1997). "The mechanism of U insertion/deletion RNA editing in kinetoplastid mitochondria". Nucleic Acids Research. 25 (19): 3751–9. doi:10.1093/nar/25.19.3751. PMC 146959. PMID 9380494.
- Blum B, Bakalara N, Simpson L (January 1990). "A model for RNA editing in kinetoplastid mitochondria: "guide" RNA molecules transcribed from maxicircle DNA provide the edited information". Cell. 60 (2): 189–98. doi:10.1016/0092-8674(90)90735-W. PMID 1688737.
- Kable ML, Heidmann S, Stuart KD (May 1997). "RNA editing: getting U into RNA". Trends in Biochemical Sciences. 22 (5): 162–6. doi:10.1016/S0968-0004(97)01041-4. PMID 9175474.
- Simpson L, Thiemann OH (June 1995). "Sense from nonsense: RNA editing in mitochondria of kinetoplastid protozoa and slime molds". Cell. 81 (6): 837–40. doi:10.1016/0092-8674(95)90003-9. PMID 7781060.
- Stuart K (February 1991). "RNA editing in mitochondrial mRNA of trypanosomatids". Trends in Biochemical Sciences. 16 (2): 68–72. doi:10.1016/0968-0004(91)90027-S. PMID 1713359.
- Hajduk SL, Sabatini RS (1998). "Mitochondrial mRNA editing in kinetoplastid protozoa". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC.: ASM Press. pp. 377–394.
- Takenaka M, Verbitskiy D, Zehrmann A, Härtel B, Bayer-Császár E, Glass F, Brennicke A (Nov 2014). "RNA editing in plant mitochondria—connecting RNA target sequences and acting proteins". Mitochondrion. Plant Mitochondria in Mitochondrion. 19 Pt B: 191–7. doi:10.1016/j.mito.2014.04.005. PMID 24732437.
- Shikanai T (September 2015). "RNA editing in plants: Machinery and flexibility of site recognition". Biochimica et Biophysica Acta. SI: Chloroplast Biogenesis. 1847 (9): 779–85. doi:10.1016/j.bbabio.2014.12.010. PMID 25585161.
- Nishikura K (2010). "Functions and regulation of RNA editing by ADAR deaminases". Annual Review of Biochemistry. 79 (1): 321–49. doi:10.1146/annurev-biochem-060208-105251. PMC 2953425. PMID 20192758.
- Tajaddod M, Jantsch MF, Licht K (March 2016). "The dynamic epitranscriptome: A to I editing modulates genetic information". Chromosoma. 125 (1): 51–63. doi:10.1007/s00412-015-0526-9. PMC 4761006. PMID 26148686.
- Licht K, Jantsch MF (April 2016). "Rapid and dynamic transcriptome regulation by RNA editing and RNA modifications". The Journal of Cell Biology. 213 (1): 15–22. doi:10.1083/jcb.201511041. PMC 4828693. PMID 27044895.
- Ramaswami G, Li JB (January 2014). "RADAR: a rigorously annotated database of A-to-I RNA editing". Nucleic Acids Research. 42 (Database issue): D109–13. doi:10.1093/nar/gkt996. PMC 3965033. PMID 24163250.
- Stulić M, Jantsch MF (October 2013). "Spatio-temporal profiling of Filamin A RNA-editing reveals ADAR preferences and high editing levels outside neuronal tissues". RNA Biology. 10 (10): 1611–7. doi:10.4161/rna.26216. PMC 3866242. PMID 24025532.
- Licht K, Kapoor U, Mayrhofer E, Jantsch MF (July 2016). "Adenosine to Inosine editing frequency controlled by splicing efficiency". Nucleic Acids Research. 44 (13): 6398–408. doi:10.1093/nar/gkw325. PMC 5291252. PMID 27112566.
- Sharma PM, Bowman M, Madden SL, Rauscher FJ, Sukumar S (March 1994). "RNA editing in the Wilms' tumor susceptibility gene, WT1". Genes & Development. 8 (6): 720–31. doi:10.1101/gad.8.6.720. PMID 7926762.
- Klimek-Tomczak K, Mikula M, Dzwonek A, Paziewska A, Karczmarski J, Hennig E, Bujnicki JM, Bragoszewski P, Denisenko O, Bomsztyk K, Ostrowski J (February 2006). "Editing of hnRNP K protein mRNA in colorectal adenocarcinoma and surrounding mucosa". British Journal of Cancer. 94 (4): 586–92. doi:10.1038/sj.bjc.6602938. PMC 2361188. PMID 16404425.
- Grohmann M, Hammer P, Walther M, Paulmann N, Büttner A, Eisenmenger W, Baghai TC, Schüle C, Rupprecht R, Bader M, Bondy B, Zill P, Priller J, Walther DJ (Jan 2010). "Alternative splicing and extensive RNA editing of human TPH2 transcripts". PLOS One. 5 (1): e8956. doi:10.1371/journal.pone.0008956. PMC 2813293. PMID 20126463.
- Castandet B, Araya A (Aug 2011). "RNA editing in plant organelles. Why make it easy?". Biochemistry. Biokhimiia. 76 (8): 924–31. doi:10.1134/S0006297911080086. PMID 22022966.
- Niavarani A, Currie E, Reyal Y, Anjos-Afonso F, Horswell S, Griessinger E, Luis Sardina J, Bonnet D (2015). "APOBEC3A is implicated in a novel class of G-to-A mRNA editing in WT1 transcripts". PLOS One. 10 (3): e0120089. doi:10.1371/journal.pone.0120089. PMC 4373805. PMID 25807502.
- Covello PS, Gray MW (October 1989). "RNA editing in plant mitochondria". Nature. 341 (6243): 662–6. doi:10.1038/341662a0. PMID 2552326.
- Gualberto JM, Lamattina L, Bonnard G, Weil JH, Grienenberger JM (October 1989). "RNA editing in wheat mitochondria results in the conservation of protein sequences". Nature. 341 (6243): 660–2. doi:10.1038/341660a0. PMID 2552325.
- Hiesel R, Wissinger B, Schuster W, Brennicke A (December 1989). "RNA editing in plant mitochondria". Science. 246 (4937): 1632–4. doi:10.1126/science.2480644. PMID 2480644.
- Hoch B, Maier RM, Appel K, Igloi GL, Kössel H (Sep 1991). "Editing of a chloroplast mRNA by creation of an initiation codon". Nature. 353 (6340): 178–80. doi:10.1038/353178a0. PMID 1653905.
- Pring D, Brennicke A, Schuster W (March 1993). "RNA editing gives a new meaning to the genetic information in mitochondria and chloroplasts". Plant Molecular Biology. 21 (6): 1163–70. doi:10.1007/BF00023611. PMID 8490134.
- Wissinger B, Brennicke A, Schuster W (September 1992). "Regenerating good sense: RNA editing and trans splicing in plant mitochondria". Trends in Genetics. 8 (9): 322–8. doi:10.1016/0168-9525(92)90265-6. PMID 1365399.
- Grienenberger, J.M. (1993). "RNA editing in plant organelles". RNA Editing (Benne, R., Ed.), Ellis Harwood, New York.
- Malek O, Lättig K, Hiesel R, Brennicke A, Knoop V (March 1996). "RNA editing in bryophytes and a molecular phylogeny of land plants". The EMBO Journal. 15 (6): 1403–11. PMC 450045. PMID 8635473.
- Freyer R, Kiefer-Meyer MC, Kössel H (June 1997). "Occurrence of plastid RNA editing in all major lineages of land plants". Proceedings of the National Academy of Sciences of the United States of America. 94 (12): 6285–90. doi:10.1073/pnas.94.12.6285. PMC 21041. PMID 9177209.
- Dietrich A, Small I, Cosset A, Weil JH, Maréchal-Drouard L (1996). "Editing and import: strategies for providing plant mitochondria with a complete set of functional transfer RNAs". Biochimie. 78 (6): 518–29. doi:10.1016/0300-9084(96)84758-4. PMID 8915541.
- Bock R, Hermann M, Fuchs M (October 1997). "Identification of critical nucleotide positions for plastid RNA editing site recognition". RNA. 3 (10): 1194–200. PMC 1369561. PMID 9326494.
- Gray MW, Covello PS (January 1993). "RNA editing in plant mitochondria and chloroplasts". FASEB Journal. 7 (1): 64–71. PMID 8422976.
- Marchfelder A, Binder S, Brennicke A, Knoop V (1998). "Preface". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC: ASM Press. pp. 307–323.
- Takenaka M, Zehrmann A, Verbitskiy D, Härtel B, Brennicke A (2013). "RNA editing in plants and its evolution". Annual Review of Genetics. 47: 335–52. doi:10.1146/annurev-genet-111212-133519. PMID 24274753.
- Barkan A, Small I (2014). "Pentatricopeptide repeat proteins in plants". Annual Review of Plant Biology. 65: 415–42. doi:10.1146/annurev-arplant-050213-040159. PMID 24471833.
- Bentolila S, Oh J, Hanson MR, Bukowski R (June 2013). "Comprehensive high-resolution analysis of the role of an Arabidopsis gene family in RNA editing". PLoS Genetics. 9 (6): e1003584. doi:10.1371/journal.pgen.1003584. PMC 3688494. PMID 23818871.
- Price DH, Gray MW (1998). "Editing of tRNA". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC: ASM Press. pp. 289–306.
- Curran J, Boeck R, Kolakofsky D (October 1991). "The Sendai virus P gene expresses both an essential protein and an inhibitor of RNA synthesis by shuffling modules via mRNA editing". The EMBO Journal. 10 (10): 3079–85. PMC 453024. PMID 1655410.
- Zheng H, Fu TB, Lazinski D, Taylor J (August 1992). "Editing on the genomic RNA of human hepatitis delta virus". Journal of Virology. 66 (8): 4693–7. PMC 241294. PMID 1629949.
- Kolakofsky D, Hausmann S (1998). "Chapter 23: Cotranscriptional Paramyxovirus mRNA Editing: a Contradiction in Terms?". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC: ASM Press. pp. 413–420.
- Carter CW (1998). "Nucleoside deaminases for cytidine and adenosine: comparisons with deaminases acting on RNA". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC: ASM Press. pp. 363–376.
- Covello PS, Gray MW (Aug 1993). "On the evolution of RNA editing". Trends in Genetics. 9 (8): 265–8. doi:10.1016/0168-9525(93)90011-6. PMID 8379005.
- Lonergan KM, Gray MW (September 1993). "Predicted editing of additional transfer RNAs in Acanthamoeba castellanii mitochondria". Nucleic Acids Research. 21 (18): 4402. doi:10.1093/nar/21.18.4402. PMC 310088. PMID 8415006.
- Bachellerie JP, Cavaille J (1998). "Small nucleolar RNAs guide the ribose methylations of eukaryotic rRNAs". In Grosjean H, Benne R. Modification and Editing of RNA. Washington, DC: ASM Press. pp. 255–272.
- Speijer D (May 2011). "Does constructive neutral evolution play an important role in the origin of cellular complexity? Making sense of the origins and uses of biological complexity". BioEssays. 33 (5): 344–9. doi:10.1002/bies.201100010. PMID 21381061.
- Witzany G (June 2011). "The agents of natural genome editing". Journal of Molecular Cell Biology. 3 (3): 181–9. doi:10.1093/jmcb/mjr005. PMID 21459884.
- Stoltzfus A (August 1999). "On the possibility of constructive neutral evolution". Journal of Molecular Evolution. 49 (2): 169–81. doi:10.1007/PL00006540. PMID 10441669.
- Agranat L, Raitskin O, Sperling J, Sperling R (April 2008). "The editing enzyme ADAR1 and the mRNA surveillance protein hUpf1 interact in the cell nucleus". Proceedings of the National Academy of Sciences of the United States of America. 105 (13): 5028–33. doi:10.1073/pnas.0710576105. PMC 2278206. PMID 18362360.