GRIA4 explained

Glutamate receptor 4 is a protein that in humans is encoded by the GRIA4 gene.^[1]

This gene is a member of a family of L-glutamate-gated ion channels that mediate fast synaptic excitatory neurotransmission. These channels are also responsive to the glutamate agonist, alpha-amino-3-hydroxy-5-methyl-4-isoxazolpropionate (AMPA). Some haplotypes of this gene show a positive association with schizophrenia. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.^[1] Like the other AMPA receptor subunits, GluA4 occurs as flip and flop spliced variant. In addition, GluA4 CTD long and short isoforms exist, and presumably an ATD-only isoform (433 aa).^[2]

Interactions

GRIA4 has been shown to interact with CACNG2,^[3] GRIP1,^[4] PICK1^[4] and PRKCG.^[5]

RNA editing

Several ion channels and neurotransmitters receptors pre-mRNa are substrates for ADARs. This includes 5 subunits of the glutamate receptor ionotropic AMPA glutamate receptor subunits (Glur2, Glur3, Glur4) and Kainate receptor subunits (Glur5, Glur6). Glutamate-gated ion channels are made up of four subunits per channel. Their function is in the mediation of fast neurotransmission to the brain. The diversity of the subunits is determined, as well as RNA splicing, by RNA editing events of the individual subunits. This give rise to the necessary diversity of the receptors. GluR4 is a gene product of the GRIA4 gene, and its pre-mRNA is subject to RNA editing.

Type

A to I RNA editing is catalyzed by a family of adenosine deaminases acting on RNA (ADARs) that specifically recognize adenosines within double-stranded regions of pre-mRNAs and deaminate them to inosine. Inosines are recognised as guanosine by the cells translational machinery. There are three members of the ADAR family ADARs 1–3, with ADAR 1 and ADAR 2 being the only enzymatically active members.ADAR3 is thought to have a regulatory role in the brain. ADAR1 and ADAR 2 are widely expressed in tissues, while ADAR 3 is restricted to the brain. The double-stranded regions of RNA are formed by base-pairing between residues in the close to region of the editing site with residues usually in a neighboring intron but can be an exonic sequence. The region that base pairs with the editing region is known as an Editing Complementary Sequence (ECS).

Location

The pre-mRNA of this subunit is edited at one position.The R/G editing site is located in exon 13 between the M3 to M4 region. Editing results in a codon change from an Arginine (AGA) to a Glycine (GGA). The location of editing corresponds to a bipartite ligand interaction domain of the receptor.((((((37))))))The R/G site is found at amino acid 769 immediately before the 3-amino-acid-long flip and flop modules introduced by alternative splicing. Flip and Flop forms are present in both edited and nonedited versions of this protein.^[6] The editing complementary sequence (ECS) is found in an intronic sequence close to the exon. The intronic sequence includes a 5' splice site, and the predicted double-stranded region is 30 base pairs in length. The adenosine residue is mismatched in genomically encoded transcript, however this is not the case following editing. Despite similar sequences to the Q/R site of GluR-B, editing this site does not occur in GluR-3 pre-mRNA. Editing results in the targeted adenosine, which is mismatched prior to editing in the double-stranded RNA structure to become matched after editing. The intronic sequence involved contains a 5' donor splice site.^{[6] [7]}

Conservation

Editing also occurs in rat.^[6]

Regulation

Editing of GluR-3 is regulated in rat brain from low levels in embryonic stage to a large increase in editing levels at birth. In humans, 80-90% of GRIA3 transcripts are edited.^[6] The absence of the Q/R site editing in this glutamate receptor subunit is due to the absence of necessary intronic sequence required to form a duplex.^[8]

Consequences

Structure

Editing results in a codon change from (AGA) to (GGA), an R to a G change at the editing site.^[6]

Function

AMPA receptors that occur in the flop form desensitise faster than the flip form. Editing at R/G site allows for faster recovery from desensitisation. Unedited Glu-R at this site have slower recovery rates. Editing, therefore, allows sustained response to rapid stimuli.

Splicing

A crosstalk between editing and splicing may occur here. Editing takes place before splicing. Like the other AMPA receptor subunits, GluA4 occurs as flip and flop spliced variant. Editing is also thought to affect splicing at this site.

See also

• AMPA receptor

Further reading

• McNamara JO, Eubanks JH, McPherson JD, Wasmuth JJ, Evans GA, Heinemann SF . Chromosomal localization of human glutamate receptor genes . The Journal of Neuroscience . 12 . 7 . 2555–2562 . July 1992 . 1319477 . 6575855 . 10.1523/JNEUROSCI.12-07-02555.1992 .
• Hardy M, Younkin D, Tang CM, Pleasure J, Shi QY, Williams M, Pleasure D . Expression of non-NMDA glutamate receptor channel genes by clonal human neurons . Journal of Neurochemistry . 63 . 2 . 482–489 . August 1994 . 7518497 . 10.1046/j.1471-4159.1994.63020482.x . 19882589 .
• Roche KW, Raymond LA, Blackstone C, Huganir RL . Transmembrane topology of the glutamate receptor subunit GluR6 . The Journal of Biological Chemistry . 269 . 16 . 11679–11682 . April 1994 . 8163463 . 10.1016/S0021-9258(17)32623-6 . free .
• Fletcher EJ, Nutt SL, Hoo KH, Elliott CE, Korczak B, McWhinnie EA, Kamboj RK . Cloning, expression and pharmacological characterization of a human glutamate receptor: hGluR4 . Receptors & Channels . 3 . 1 . 21–31 . 1996 . 8589990 .
• Bonaldo MF, Lennon G, Soares MB . Normalization and subtraction: two approaches to facilitate gene discovery . Genome Research . 6 . 9 . 791–806 . September 1996 . 8889548 . 10.1101/gr.6.9.791 . free .
• Ripellino JA, Neve RL, Howe JR . Expression and heteromeric interactions of non-N-methyl-D-aspartate glutamate receptor subunits in the developing and adult cerebellum . Neuroscience . 82 . 2 . 485–497 . January 1998 . 9466455 . 10.1016/S0306-4522(97)00296-0 . 23219004 . free .
• Carvalho AL, Kameyama K, Huganir RL . Characterization of phosphorylation sites on the glutamate receptor 4 subunit of the AMPA receptors . The Journal of Neuroscience . 19 . 12 . 4748–4754 . June 1999 . 10366608 . 6782640 . 10.1523/JNEUROSCI.19-12-04748.1999 .
• Chen L, Chetkovich DM, Petralia RS, Sweeney NT, Kawasaki Y, Wenthold RJ, Bredt DS, Nicoll RA . Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms . Nature . 408 . 6815 . 936–943 . 2001 . 11140673 . 10.1038/35050030 . 2000Natur.408..936C . 4427689 .
• Hirbec H, Perestenko O, Nishimune A, Meyer G, Nakanishi S, Henley JM, Dev KK . The PDZ proteins PICK1, GRIP, and syntenin bind multiple glutamate receptor subtypes. Analysis of PDZ binding motifs . The Journal of Biological Chemistry . 277 . 18 . 15221–15224 . May 2002 . 11891216 . 10.1074/jbc.C200112200 . free . 2262/89271 . free .
• Tomiyama M, Rodríguez-Puertas R, Cortés R, Pazos A, Palacios JM, Mengod G . Flip and flop splice variants of AMPA receptor subunits in the spinal cord of amyotrophic lateral sclerosis . Synapse . 45 . 4 . 245–249 . September 2002 . 12125045 . 10.1002/syn.10098 . 28604714 . 10.1.1.575.9300 .
• Pasternack A, Coleman SK, Jouppila A, Mottershead DG, Lindfors M, Pasternack M, Keinänen K . Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor channels lacking the N-terminal domain . The Journal of Biological Chemistry . 277 . 51 . 49662–49667 . December 2002 . 12393905 . 10.1074/jbc.M208349200 . free .
• Correia SS, Duarte CB, Faro CJ, Pires EV, Carvalho AL . Protein kinase C gamma associates directly with the GluR4 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor subunit. Effect on receptor phosphorylation . The Journal of Biological Chemistry . 278 . 8 . 6307–6313 . February 2003 . 12471040 . 10.1074/jbc.M205587200 . free . 10316/12633 . free .
• Makino C, Fujii Y, Kikuta R, Hirata N, Tani A, Shibata A, Ninomiya H, Tashiro N, Shibata H, Fukumaki Y . Positive association of the AMPA receptor subunit GluR4 gene (GRIA4) haplotype with schizophrenia: linkage disequilibrium mapping using SNPs evenly distributed across the gene region . American Journal of Medical Genetics. Part B, Neuropsychiatric Genetics . 116B . 1 . 17–22 . January 2003 . 12497607 . 10.1002/ajmg.b.10041 . 24883590 .
• Coleman SK, Cai C, Mottershead DG, Haapalahti JP, Keinänen K . Surface expression of GluR-D AMPA receptor is dependent on an interaction between its C-terminal domain and a 4.1 protein . The Journal of Neuroscience . 23 . 3 . 798–806 . February 2003 . 12574408 . 6741938 . 10.1523/JNEUROSCI.23-03-00798.2003 .
• Pasternack A, Coleman SK, Féthière J, Madden DR, LeCaer JP, Rossier J, Pasternack M, Keinänen K . Characterization of the functional role of the N-glycans in the AMPA receptor ligand-binding domain . Journal of Neurochemistry . 84 . 5 . 1184–1192 . March 2003 . 12603841 . 10.1046/j.1471-4159.2003.01611.x . 11350496 . free .
• Kawahara Y, Ito K, Sun H, Ito M, Kanazawa I, Kwak S . GluR4c, an alternative splicing isoform of GluR4, is abundantly expressed in the adult human brain . Brain Research. Molecular Brain Research . 127 . 1–2 . 150–155 . August 2004 . 15306133 . 10.1016/j.molbrainres.2004.05.020 .
• Li G, Sheng Z, Huang Z, Niu L . Kinetic mechanism of channel opening of the GluRDflip AMPA receptor . Biochemistry . 44 . 15 . 5835–5841 . April 2005 . 15823042 . 10.1021/bi047413n .
• Nuriya M, Oh S, Huganir RL . Phosphorylation-dependent interactions of alpha-Actinin-1/IQGAP1 with the AMPA receptor subunit GluR4 . Journal of Neurochemistry . 95 . 2 . 544–552 . October 2005 . 16190873 . 10.1111/j.1471-4159.2005.03410.x . 9517535 .

Notes and References

1. Web site: Entrez Gene: GRIA4 glutamate receptor, ionotrophic, AMPA 4.
2. Herbrechter R, Hube N, Buchholz R, Reiner A . Splicing and editing of ionotropic glutamate receptors: a comprehensive analysis based on human RNA-Seq data . Cellular and Molecular Life Sciences . 78 . 14 . 5605–5630 . July 2021 . 34100982 . 10.1007/s00018-021-03865-z. 8257547 . 1420-682X .
3. Chen L, Chetkovich DM, Petralia RS, Sweeney NT, Kawasaki Y, Wenthold RJ, Bredt DS, Nicoll RA . Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms . Nature . 408 . 6815 . 936–943 . 2000 . 11140673 . 10.1038/35050030 . 4427689 . 2000Natur.408..936C .
4. Hirbec H, Perestenko O, Nishimune A, Meyer G, Nakanishi S, Henley JM, Dev KK . The PDZ proteins PICK1, GRIP, and syntenin bind multiple glutamate receptor subtypes. Analysis of PDZ binding motifs . The Journal of Biological Chemistry . 277 . 18 . 15221–15224 . May 2002 . 11891216 . 10.1074/jbc.C200112200 . free . 2262/89271 . free .
5. Correia SS, Duarte CB, Faro CJ, Pires EV, Carvalho AL . Protein kinase C gamma associates directly with the GluR4 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor subunit. Effect on receptor phosphorylation . The Journal of Biological Chemistry . 278 . 8 . 6307–6313 . February 2003 . 12471040 . 10.1074/jbc.M205587200 . free . 10316/12633 . free .
6. Lomeli H, Mosbacher J, Melcher T, Höger T, Geiger JR, Kuner T, Monyer H, Higuchi M, Bach A, Seeburg PH . Control of kinetic properties of AMPA receptor channels by nuclear RNA editing . Science . 266 . 5191 . 1709–1713 . December 1994 . 7992055 . 10.1126/science.7992055 . 1994Sci...266.1709L .
7. Seeburg PH, Higuchi M, Sprengel R . RNA editing of brain glutamate receptor channels: mechanism and physiology . Brain Research. Brain Research Reviews . 26 . 2–3 . 217–229 . May 1998 . 9651532 . 10.1016/S0165-0173(97)00062-3 . 12147763 .
8. Herb A, Higuchi M, Sprengel R, Seeburg PH . Q/R site editing in kainate receptor GluR5 and GluR6 pre-mRNAs requires distant intronic sequences . Proceedings of the National Academy of Sciences of the United States of America . 93 . 5 . 1875–1880 . March 1996 . 8700852 . 39875 . 10.1073/pnas.93.5.1875 . free . 1996PNAS...93.1875H .