Abstract
Go alpha a guanine nucleotide-binding (G) protein abundant in brain and other neural tissues, has been implicated in ion channel regulation. Concerted efforts in several laboratories have revealed multiple Go alpha mRNAs and protein isoforms in different contexts. Go alpha is a single copy gene in mammalian species, although the structure, number and tissue localization of Go alpha mRNAs reported by investigators are inconsistent. To define the cell-specific expression of alternatively spliced variants of Go alpha mRNA, we employed several strategies, including Northern hybridizations with sequences-specific oligonucleotides, selective digestions of Go alpha mRNA using RNase H, and adaptations of the polymerase chain reaction. Four distinct alternatively spliced variants were identified, a 5.7-kb Go alpha 2 mRNA and three Go alpha 1 mRNAs with different 3' UTRs. The UTRs of the three Go alpha 1s are composed of different combinations of what have been referred to as UTR-A and UTR-B. The sequences of the spliced segments are well conserved among mammalian species, suggesting a functional role for these alternatively spliced 3' UTRs in post-transcriptional and/or tissue-specific regulation of Go alpha expression. The position of the intron-exon splice boundary at nucleotide 31 following T of the TGA stop codon is conserved in the Gi alpha 2 and Gi alpha 3 genes, consistent with the notion that similar alternative splicing of 3' UTRs occurs in products of these related genes.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aziz N., Munro H. N. Iron regulates ferritin mRNA translation through a segment of its 5' untranslated region. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8478–8482. doi: 10.1073/pnas.84.23.8478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berger S. L. Direct mapping of rare messenger RNAs by means of oligomer-directed ribonuclease H cleavage. Anal Biochem. 1987 Mar;161(2):272–279. doi: 10.1016/0003-2697(87)90451-9. [DOI] [PubMed] [Google Scholar]
- Bertrand P., Sanford J., Rudolph U., Codina J., Birnbaumer L. At least three alternatively spliced mRNAs encoding two alpha subunits of the Go GTP-binding protein can be expressed in a single tissue. J Biol Chem. 1990 Oct 25;265(30):18576–18580. [PubMed] [Google Scholar]
- Birnbaumer L. Transduction of receptor signal into modulation of effector activity by G proteins: the first 20 years or so .... FASEB J. 1990 Nov;4(14):3178–3188. doi: 10.1096/fasebj.4.14.2172060. [DOI] [PubMed] [Google Scholar]
- Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
- Brabet P., Dumuis A., Sebben M., Pantaloni C., Bockaert J., Homburger V. Immunocytochemical localization of the guanine nucleotide-binding protein Go in primary cultures of neuronal and glial cells. J Neurosci. 1988 Feb;8(2):701–708. doi: 10.1523/JNEUROSCI.08-02-00701.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brabet P., Pantaloni C., Bockaert J., Homburger V. Metabolism of two Go alpha isoforms in neuronal cells during differentiation. J Biol Chem. 1991 Jul 15;266(20):12825–12828. [PubMed] [Google Scholar]
- Brabet P., Pantaloni C., Rodriguez M., Martinez J., Bockaert J., Homburger V. Neuroblastoma differentiation involves the expression of two isoforms of the alpha-subunit of Go. J Neurochem. 1990 Apr;54(4):1310–1320. doi: 10.1111/j.1471-4159.1990.tb01964.x. [DOI] [PubMed] [Google Scholar]
- Brann M. R., Collins R. M., Spiegel A. Localization of mRNAs encoding the alpha-subunits of signal-transducing G-proteins within rat brain and among peripheral tissues. FEBS Lett. 1987 Sep 28;222(1):191–198. doi: 10.1016/0014-5793(87)80218-1. [DOI] [PubMed] [Google Scholar]
- Brawerman G. Determinants of messenger RNA stability. Cell. 1987 Jan 16;48(1):5–6. doi: 10.1016/0092-8674(87)90346-1. [DOI] [PubMed] [Google Scholar]
- Bray P., Carter A., Guo V., Puckett C., Kamholz J., Spiegel A., Nirenberg M. Human cDNA clones for an alpha subunit of Gi signal-transduction protein. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5115–5119. doi: 10.1073/pnas.84.15.5115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casey P. J., Gilman A. G. G protein involvement in receptor-effector coupling. J Biol Chem. 1988 Feb 25;263(6):2577–2580. [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Eisenstein R. S., Munro H. N. Translational regulation of ferritin synthesis by iron. Enzyme. 1990;44(1-4):42–58. doi: 10.1159/000468746. [DOI] [PubMed] [Google Scholar]
- Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gierschik P., Morrow B., Milligan G., Rubin C., Spiegel A. Changes in the guanine nucleotide-binding proteins, Gi and Go, during differentiation of 3T3-L1 cells. FEBS Lett. 1986 Apr 7;199(1):103–106. doi: 10.1016/0014-5793(86)81233-9. [DOI] [PubMed] [Google Scholar]
- Gillis P., Malter J. S. The adenosine-uridine binding factor recognizes the AU-rich elements of cytokine, lymphokine, and oncogene mRNAs. J Biol Chem. 1991 Feb 15;266(5):3172–3177. [PubMed] [Google Scholar]
- Goldsmith P., Backlund P. S., Jr, Rossiter K., Carter A., Milligan G., Unson C. G., Spiegel A. Purification of heterotrimeric GTP-binding proteins from brain: identification of a novel form of Go. Biochemistry. 1988 Sep 6;27(18):7085–7090. doi: 10.1021/bi00418a062. [DOI] [PubMed] [Google Scholar]
- Hescheler J., Rosenthal W., Trautwein W., Schultz G. The GTP-binding protein, Go, regulates neuronal calcium channels. 1987 Jan 29-Feb 4Nature. 325(6103):445–447. doi: 10.1038/325445a0. [DOI] [PubMed] [Google Scholar]
- Hsu W. H., Rudolph U., Sanford J., Bertrand P., Olate J., Nelson C., Moss L. G., Boyd A. E., Codina J., Birnbaumer L. Molecular cloning of a novel splice variant of the alpha subunit of the mammalian Go protein. J Biol Chem. 1990 Jul 5;265(19):11220–11226. [PubMed] [Google Scholar]
- Hultman T., Bergh S., Moks T., Uhlén M. Bidirectional solid-phase sequencing of in vitro-amplified plasmid DNA. Biotechniques. 1991 Jan;10(1):84–93. [PubMed] [Google Scholar]
- Itoh H., Kozasa T., Nagata S., Nakamura S., Katada T., Ui M., Iwai S., Ohtsuka E., Kawasaki H., Suzuki K. Molecular cloning and sequence determination of cDNAs for alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go from rat brain. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3776–3780. doi: 10.1073/pnas.83.11.3776. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itoh H., Toyama R., Kozasa T., Tsukamoto T., Matsuoka M., Kaziro Y. Presence of three distinct molecular species of Gi protein alpha subunit. Structure of rat cDNAs and human genomic DNAs. J Biol Chem. 1988 May 15;263(14):6656–6664. [PubMed] [Google Scholar]
- Jackson R. J., Standart N. Do the poly(A) tail and 3' untranslated region control mRNA translation? Cell. 1990 Jul 13;62(1):15–24. doi: 10.1016/0092-8674(90)90235-7. [DOI] [PubMed] [Google Scholar]
- Jones D. T., Reed R. R. Molecular cloning of five GTP-binding protein cDNA species from rat olfactory neuroepithelium. J Biol Chem. 1987 Oct 15;262(29):14241–14249. [PubMed] [Google Scholar]
- Kim S. J., Park K., Koeller D., Kim K. Y., Wakefield L. M., Sporn M. B., Roberts A. B. Post-transcriptional regulation of the human transforming growth factor-beta 1 gene. J Biol Chem. 1992 Jul 5;267(19):13702–13707. [PubMed] [Google Scholar]
- Kobayashi I., Shibasaki H., Takahashi K., Kikkawa S., Ui M., Katada T. Purification of GTP-binding proteins from bovine brain membranes. Identification of heterogeneity of the alpha-subunit of Go proteins. FEBS Lett. 1989 Oct 23;257(1):177–180. doi: 10.1016/0014-5793(89)81815-0. [DOI] [PubMed] [Google Scholar]
- Kobayashi I., Shibasaki H., Takahashi K., Tohyama K., Kurachi Y., Ito H., Ui M., Katada T. Purification and characterization of five different alpha subunits of guanine-nucleotide-binding proteins in bovine brain membranes. Their physiological properties concerning the activities of adenylate cyclase and atrial muscarinic K+ channels. Eur J Biochem. 1990 Jul 31;191(2):499–506. doi: 10.1111/j.1432-1033.1990.tb19149.x. [DOI] [PubMed] [Google Scholar]
- Kozasa T., Itoh H., Tsukamoto T., Kaziro Y. Isolation and characterization of the human Gs alpha gene. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2081–2085. doi: 10.1073/pnas.85.7.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lavu S., Clark J., Swarup R., Matsushima K., Paturu K., Moss J., Kung H. F. Molecular cloning and DNA sequence analysis of the human guanine nucleotide-binding protein Go alpha. Biochem Biophys Res Commun. 1988 Jan 29;150(2):811–815. doi: 10.1016/0006-291x(88)90463-9. [DOI] [PubMed] [Google Scholar]
- Lee W. M., Lin C., Curran T. Activation of the transforming potential of the human fos proto-oncogene requires message stabilization and results in increased amounts of partially modified fos protein. Mol Cell Biol. 1988 Dec;8(12):5521–5527. doi: 10.1128/mcb.8.12.5521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liang B. T., Hellmich M. R., Neer E. J., Galper J. B. Development of muscarinic cholinergic inhibition of adenylate cyclase in embryonic chick heart. Its relationship to changes in the inhibitory guanine nucleotide regulatory protein. J Biol Chem. 1986 Jul 5;261(19):9011–9021. [PubMed] [Google Scholar]
- Luetje C. W., Nathanson N. M. Chronic membrane depolarization regulates the level of the guanine nucleotide binding protein Go alpha in cultured neuronal cells. J Neurochem. 1988 Jun;50(6):1775–1782. doi: 10.1111/j.1471-4159.1988.tb02478.x. [DOI] [PubMed] [Google Scholar]
- Luetje C. W., Tietje K. M., Christian J. L., Nathanson N. M. Differential tissue expression and developmental regulation of guanine nucleotide binding regulatory proteins and their messenger RNAs in rat heart. J Biol Chem. 1988 Sep 15;263(26):13357–13365. [PubMed] [Google Scholar]
- Milligan G., Tanfin Z., Goureau O., Unson C., Harbon S. Identification of both Gi2 and a novel, immunologically distinct, form of Go in rat myometrial membranes. FEBS Lett. 1989 Feb 27;244(2):411–416. doi: 10.1016/0014-5793(89)80574-5. [DOI] [PubMed] [Google Scholar]
- Mitchell L. G., Merril C. R. Affinity generation of single-stranded DNA for dideoxy sequencing following the polymerase chain reaction. Anal Biochem. 1989 May 1;178(2):239–242. doi: 10.1016/0003-2697(89)90631-3. [DOI] [PubMed] [Google Scholar]
- Mullaney I., Magee A. I., Unson C. G., Milligan G. Differential regulation of amounts of the guanine-nucleotide-binding proteins Gi and Go in neuroblastoma x glioma hybrid cells in response to dibutyryl cyclic AMP. Biochem J. 1988 Dec 1;256(2):649–656. doi: 10.1042/bj2560649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murtagh J. J., Jr, Eddy R., Shows T. B., Moss J., Vaughan M. Different forms of Go alpha mRNA arise by alternative splicing of transcripts from a single gene on human chromosome 16. Mol Cell Biol. 1991 Feb;11(2):1146–1155. doi: 10.1128/mcb.11.2.1146. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neer E. J., Lok J. M., Wolf L. G. Purification and properties of the inhibitory guanine nucleotide regulatory unit of brain adenylate cyclase. J Biol Chem. 1984 Nov 25;259(22):14222–14229. [PubMed] [Google Scholar]
- Nestler E. J., Terwilliger R. Z., Walker J. R., Sevarino K. A., Duman R. S. Chronic cocaine treatment decreases levels of the G protein subunits Gi alpha and Go alpha in discrete regions of rat brain. J Neurochem. 1990 Sep;55(3):1079–1082. doi: 10.1111/j.1471-4159.1990.tb04602.x. [DOI] [PubMed] [Google Scholar]
- Price S. R., Murtagh J. J., Jr, Tsuchiya M., Serventi I. M., Van Meurs K. P., Angus C. W., Moss J., Vaughan M. Multiple forms of Go alpha mRNA: analysis of the 3'-untranslated regions. Biochemistry. 1990 May 29;29(21):5069–5076. doi: 10.1021/bi00473a011. [DOI] [PubMed] [Google Scholar]
- Price S. R., Tsai S. C., Adamik R., Angus C. W., Serventi I. M., Tsuchiya M., Moss J., Vaughan M. Expression of Go alpha mRNA and protein in bovine tissues. Biochemistry. 1989 May 2;28(9):3803–3807. doi: 10.1021/bi00435a027. [DOI] [PubMed] [Google Scholar]
- Rapiejko P. J., Watkins D. C., Ros M., Malbon C. C. G-protein subunit mRNA levels in rat heart, liver, and adipose tissues: analysis by DNA-excess solution hybridization. Biochim Biophys Acta. 1990 May 2;1052(2):348–350. doi: 10.1016/0167-4889(90)90233-4. [DOI] [PubMed] [Google Scholar]
- Raymond V., Atwater J. A., Verma I. M. Removal of an mRNA destabilizing element correlates with the increased oncogenicity of proto-oncogene fos. Oncogene Res. 1989;5(1):1–12. [PubMed] [Google Scholar]
- Rosenthal A., Jones D. S. Genomic walking and sequencing by oligo-cassette mediated polymerase chain reaction. Nucleic Acids Res. 1990 May 25;18(10):3095–3096. doi: 10.1093/nar/18.10.3095. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rouot B., Carrette J., Lafontan M., Lan Tran P., Fehrentz J. A., Bockaert J., Toutant M. The adipocyte Go alpha-immunoreactive polypeptide is different from the alpha subunit of the brain Go protein. Biochem J. 1989 May 15;260(1):307–310. doi: 10.1042/bj2600307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rouot B., Charpentier N., Chabbert C., Carrette J., Zumbihl R., Bockaert J., Homburger V. Specific antibodies against Go isoforms reveal the early expression of the Go2 alpha subunit and appearance of Go1 alpha during neuronal differentiation. Mol Pharmacol. 1992 Feb;41(2):273–280. [PubMed] [Google Scholar]
- Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
- Shibasaki H., Kozasa T., Takahashi K., Inanobe A., Kaziro Y., Ui M., Katada T. Amino acid sequence determination of the novel forms of Go alpha purified from bovine brain membranes. FEBS Lett. 1991 Jul 22;285(2):268–270. doi: 10.1016/0014-5793(91)80814-j. [DOI] [PubMed] [Google Scholar]
- Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
- Sternweis P. C., Robishaw J. D. Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem. 1984 Nov 25;259(22):13806–13813. [PubMed] [Google Scholar]
- Strathmann M., Wilkie T. M., Simon M. I. Alternative splicing produces transcripts encoding two forms of the alpha subunit of GTP-binding protein Go. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6477–6481. doi: 10.1073/pnas.87.17.6477. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strittmatter S. M., Valenzuela D., Kennedy T. E., Neer E. J., Fishman M. C. G0 is a major growth cone protein subject to regulation by GAP-43. Nature. 1990 Apr 26;344(6269):836–841. doi: 10.1038/344836a0. [DOI] [PubMed] [Google Scholar]
- Tsuchiya M., Price S. R., Nightingale M. S., Moss J., Vaughan M. Tissue and species distribution of mRNA encoding two ADP-ribosylation factors, 20-kDa guanine nucleotide binding proteins. Biochemistry. 1989 Dec 12;28(25):9668–9673. doi: 10.1021/bi00451a019. [DOI] [PubMed] [Google Scholar]
- Tsukamoto T., Toyama R., Itoh H., Kozasa T., Matsuoka M., Kaziro Y. Structure of the human gene and two rat cDNAs encoding the alpha chain of GTP-binding regulatory protein Go: two different mRNAs are generated by alternative splicing. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):2974–2978. doi: 10.1073/pnas.88.8.2974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Meurs K. P., Angus C. W., Lavu S., Kung H. F., Czarnecki S. K., Moss J., Vaughan M. Deduced amino acid sequence of bovine retinal Go alpha: similarities to other guanine nucleotide-binding proteins. Proc Natl Acad Sci U S A. 1987 May;84(10):3107–3111. doi: 10.1073/pnas.84.10.3107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watkins D. C., Northup J. K., Malbon C. C. Regulation of G-proteins in differentiation. Altered ratio of alpha- to beta-subunits in 3T3-L1 cells. J Biol Chem. 1987 Aug 5;262(22):10651–10657. [PubMed] [Google Scholar]
- Worley P. F., Baraban J. M., Van Dop C., Neer E. J., Snyder S. H. Go, a guanine nucleotide-binding protein: immunohistochemical localization in rat brain resembles distribution of second messenger systems. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4561–4565. doi: 10.1073/pnas.83.12.4561. [DOI] [PMC free article] [PubMed] [Google Scholar]


