Abstract
Ubiquitin is a multifunctional 76-amino-acid protein which plays critical roles in many aspects of cellular metabolism. In Caenorhabditis elegans, the major source of ubiquitin RNA is the polyubiquitin locus, UbiA. UbiA is transcribed as a polycistronic mRNA which contains 11 tandem repeats of ubiquitin sequence and possesses a 2-amino-acid carboxy-terminal extension on the final repeat. The UbiA locus possesses several unusual features not seen in the ubiquitin genes of other organisms studied to date. Mature UbiA mRNA acquires a 22-nucleotide leader sequence via a trans-splicing reaction involving a 100-nucleotide splice leader RNA derived from a different chromosome. UbiA is also unique among known polyubiquitin genes in containing four cis-spliced introns within its coding sequence. Thus, UbiA is one of a small class of genes found in higher eucaryotes whose heterogeneous nuclear RNA undergoes both cis and trans splicing. The putative promoter region of UbiA contains a number of potential regulatory elements: (i) a cytosine-rich block, (ii) two sequences resembling the heat shock regulatory element, and (iii) a palindromic sequence with homology to the DNA-binding site of the mammalian steroid hormone receptor. The expression of the UbiA gene has been studied under various heat shock conditions and has been monitored during larval moulting and throughout the major stages of development. These studies indicate that the expression of the UbiA gene is not inducible by acute or chronic heat shock and does not appear to be under nutritional or developmental regulation in C. elegans.
Full text
PDF![268](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/e0f160e0876d/molcellb00049-0284.png)
![269](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/58df3418f424/molcellb00049-0285.png)
![270](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/b352318e8d05/molcellb00049-0286.png)
![271](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/1bdfc6414391/molcellb00049-0287.png)
![272](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/bfbdc6a37c05/molcellb00049-0288.png)
![273](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/cb7a9fe0f4ba/molcellb00049-0289.png)
![274](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/855c9b6c61a0/molcellb00049-0290.png)
![275](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/012e544fc5c1/molcellb00049-0291.png)
![276](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/29702f310efa/molcellb00049-0292.png)
![277](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ae0/362169/c44f5daea14b/molcellb00049-0293.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ananthan J., Goldberg A. L., Voellmy R. Abnormal proteins serve as eukaryotic stress signals and trigger the activation of heat shock genes. Science. 1986 Apr 25;232(4749):522–524. doi: 10.1126/science.3083508. [DOI] [PubMed] [Google Scholar]
- Baker R. T., Board P. G. The human ubiquitin gene family: structure of a gene and pseudogenes from the Ub B subfamily. Nucleic Acids Res. 1987 Jan 26;15(2):443–463. doi: 10.1093/nar/15.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ball E., Karlik C. C., Beall C. J., Saville D. L., Sparrow J. C., Bullard B., Fyrberg E. A. Arthrin, a myofibrillar protein of insect flight muscle, is an actin-ubiquitin conjugate. Cell. 1987 Oct 23;51(2):221–228. doi: 10.1016/0092-8674(87)90149-8. [DOI] [PubMed] [Google Scholar]
- Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
- Bond U., Schlesinger M. J. Ubiquitin is a heat shock protein in chicken embryo fibroblasts. Mol Cell Biol. 1985 May;5(5):949–956. doi: 10.1128/mcb.5.5.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burdon R. H. Heat shock and the heat shock proteins. Biochem J. 1986 Dec 1;240(2):313–324. doi: 10.1042/bj2400313. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cassada R. C., Russell R. L. The dauerlarva, a post-embryonic developmental variant of the nematode Caenorhabditis elegans. Dev Biol. 1975 Oct;46(2):326–342. doi: 10.1016/0012-1606(75)90109-8. [DOI] [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]
- Ciechanover A., Finley D., Varshavsky A. Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant ts85. Cell. 1984 May;37(1):57–66. doi: 10.1016/0092-8674(84)90300-3. [DOI] [PubMed] [Google Scholar]
- Coulson A., Sulston J., Brenner S., Karn J. Toward a physical map of the genome of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821–7825. doi: 10.1073/pnas.83.20.7821. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Craig E. A. The heat shock response. CRC Crit Rev Biochem. 1985;18(3):239–280. doi: 10.3109/10409238509085135. [DOI] [PubMed] [Google Scholar]
- Delaney A. D. A DNA sequence handling program. Nucleic Acids Res. 1982 Jan 11;10(1):61–67. doi: 10.1093/nar/10.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Douglass J., Civelli O., Herbert E. Polyprotein gene expression: generation of diversity of neuroendocrine peptides. Annu Rev Biochem. 1984;53:665–715. doi: 10.1146/annurev.bi.53.070184.003313. [DOI] [PubMed] [Google Scholar]
- Dworkin-Rastl E., Shrutkowski A., Dworkin M. B. Multiple ubiquitin mRNAs during Xenopus laevis development contain tandem repeats of the 76 amino acid coding sequence. Cell. 1984 Dec;39(2 Pt 1):321–325. doi: 10.1016/0092-8674(84)90010-2. [DOI] [PubMed] [Google Scholar]
- Emmons S. W., Klass M. R., Hirsh D. Analysis of the constancy of DNA sequences during development and evolution of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1333–1337. doi: 10.1073/pnas.76.3.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finley D., Ciechanover A., Varshavsky A. Thermolability of ubiquitin-activating enzyme from the mammalian cell cycle mutant ts85. Cell. 1984 May;37(1):43–55. doi: 10.1016/0092-8674(84)90299-x. [DOI] [PubMed] [Google Scholar]
- Finley D., Ozkaynak E., Varshavsky A. The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. Cell. 1987 Mar 27;48(6):1035–1046. doi: 10.1016/0092-8674(87)90711-2. [DOI] [PubMed] [Google Scholar]
- Fried V. A., Smith H. T., Hildebrandt E., Weiner K. Ubiquitin has intrinsic proteolytic activity: implications for cellular regulation. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3685–3689. doi: 10.1073/pnas.84.11.3685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gausing K., Barkardottir R. Structure and expression of ubiquitin genes in higher plants. Eur J Biochem. 1986 Jul 1;158(1):57–62. doi: 10.1111/j.1432-1033.1986.tb09720.x. [DOI] [PubMed] [Google Scholar]
- Giorda R., Ennis H. L. Structure of two developmentally regulated Dictyostelium discoideum ubiquitin genes. Mol Cell Biol. 1987 Jun;7(6):2097–2103. doi: 10.1128/mcb.7.6.2097. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goff S. A., Goldberg A. L. Production of abnormal proteins in E. coli stimulates transcription of lon and other heat shock genes. Cell. 1985 Jun;41(2):587–595. doi: 10.1016/s0092-8674(85)80031-3. [DOI] [PubMed] [Google Scholar]
- Graham R. W., Van Doren K., Bektesh S., Candido E. P. Maturation of the major ubiquitin gene transcript in Caenorhabditis elegans involves the acquisition of a trans-spliced leader. J Biol Chem. 1988 Jul 25;263(21):10415–10419. [PubMed] [Google Scholar]
- Hershko A., Ciechanover A. The ubiquitin pathway for the degradation of intracellular proteins. Prog Nucleic Acid Res Mol Biol. 1986;33:19-56, 301. doi: 10.1016/s0079-6603(08)60019-7. [DOI] [PubMed] [Google Scholar]
- Hiromi Y., Okamoto H., Gehring W. J., Hotta Y. Germline transformation with Drosophila mutant actin genes induces constitutive expression of heat shock genes. Cell. 1986 Jan 31;44(2):293–301. doi: 10.1016/0092-8674(86)90763-4. [DOI] [PubMed] [Google Scholar]
- Jahngen J. H., Haas A. L., Ciechanover A., Blondin J., Eisenhauer D., Taylor A. The eye lens has an active ubiquitin-protein conjugation system. J Biol Chem. 1986 Oct 15;261(29):13760–13767. [PubMed] [Google Scholar]
- Jentsch S., McGrath J. P., Varshavsky A. The yeast DNA repair gene RAD6 encodes a ubiquitin-conjugating enzyme. Nature. 1987 Sep 10;329(6135):131–134. doi: 10.1038/329131a0. [DOI] [PubMed] [Google Scholar]
- Jones D., Russnak R. H., Kay R. J., Candido E. P. Structure, expression, and evolution of a heat shock gene locus in Caenorhabditis elegans that is flanked by repetitive elements. J Biol Chem. 1986 Sep 15;261(26):12006–12015. [PubMed] [Google Scholar]
- Jones K. A., Yamamoto K. R., Tjian R. Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro. Cell. 1985 Sep;42(2):559–572. doi: 10.1016/0092-8674(85)90113-8. [DOI] [PubMed] [Google Scholar]
- Krause M., Hirsh D. A trans-spliced leader sequence on actin mRNA in C. elegans. Cell. 1987 Jun 19;49(6):753–761. doi: 10.1016/0092-8674(87)90613-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurjan J., Herskowitz I. Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor. Cell. 1982 Oct;30(3):933–943. doi: 10.1016/0092-8674(82)90298-7. [DOI] [PubMed] [Google Scholar]
- Leung D. W., Spencer S. A., Cachianes G., Hammonds R. G., Collins C., Henzel W. J., Barnard R., Waters M. J., Wood W. I. Growth hormone receptor and serum binding protein: purification, cloning and expression. Nature. 1987 Dec 10;330(6148):537–543. doi: 10.1038/330537a0. [DOI] [PubMed] [Google Scholar]
- Levinger L., Varshavsky A. Selective arrangement of ubiquitinated and D1 protein-containing nucleosomes within the Drosophila genome. Cell. 1982 Feb;28(2):375–385. doi: 10.1016/0092-8674(82)90355-5. [DOI] [PubMed] [Google Scholar]
- Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
- McGhee J. D., Wood W. I., Dolan M., Engel J. D., Felsenfeld G. A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion. Cell. 1981 Nov;27(1 Pt 2):45–55. doi: 10.1016/0092-8674(81)90359-7. [DOI] [PubMed] [Google Scholar]
- McLauchlan J., Gaffney D., Whitton J. L., Clements J. B. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini. Nucleic Acids Res. 1985 Feb 25;13(4):1347–1368. doi: 10.1093/nar/13.4.1347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
- Mezquita J., Oliva R., Mezquita C. New ubiquitin mRNA expressed during chicken spermiogenesis. Nucleic Acids Res. 1987 Nov 25;15(22):9604–9604. doi: 10.1093/nar/15.22.9604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Munro S., Pelham H. What turns on heat shock genes? Nature. 1985 Oct 10;317(6037):477–478. doi: 10.1038/317477a0. [DOI] [PubMed] [Google Scholar]
- Nevins J. R. The pathway of eukaryotic mRNA formation. Annu Rev Biochem. 1983;52:441–466. doi: 10.1146/annurev.bi.52.070183.002301. [DOI] [PubMed] [Google Scholar]
- Ozkaynak E., Finley D., Solomon M. J., Varshavsky A. The yeast ubiquitin genes: a family of natural gene fusions. EMBO J. 1987 May;6(5):1429–1439. doi: 10.1002/j.1460-2075.1987.tb02384.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ozkaynak E., Finley D., Varshavsky A. The yeast ubiquitin gene: head-to-tail repeats encoding a polyubiquitin precursor protein. Nature. 1984 Dec 13;312(5995):663–666. doi: 10.1038/312663a0. [DOI] [PubMed] [Google Scholar]
- Parag H. A., Raboy B., Kulka R. G. Effect of heat shock on protein degradation in mammalian cells: involvement of the ubiquitin system. EMBO J. 1987 Jan;6(1):55–61. doi: 10.1002/j.1460-2075.1987.tb04718.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
- Pickart C. M., Rose I. A. Functional heterogeneity of ubiquitin carrier proteins. J Biol Chem. 1985 Feb 10;260(3):1573–1581. [PubMed] [Google Scholar]
- Rapoport S. M., Dubiel W., Müller M. The mechanism of maturation-dependent breakdown of mitochondria in reticulocytes. Acta Biol Med Ger. 1981;40(10-11):1277–1283. [PubMed] [Google Scholar]
- Rechsteiner M. Ubiquitin-mediated pathways for intracellular proteolysis. Annu Rev Cell Biol. 1987;3:1–30. doi: 10.1146/annurev.cb.03.110187.000245. [DOI] [PubMed] [Google Scholar]
- Rosenzweig B., Liao L. W., Hirsh D. Sequence of the C. elegans transposable element Tc1. Nucleic Acids Res. 1983 Jun 25;11(12):4201–4209. doi: 10.1093/nar/11.12.4201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Russnak R. H., Candido E. P. Locus encoding a family of small heat shock genes in Caenorhabditis elegans: two genes duplicated to form a 3.8-kilobase inverted repeat. Mol Cell Biol. 1985 Jun;5(6):1268–1278. doi: 10.1128/mcb.5.6.1268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scheidereit C., Geisse S., Westphal H. M., Beato M. The glucocorticoid receptor binds to defined nucleotide sequences near the promoter of mouse mammary tumour virus. Nature. 1983 Aug 25;304(5928):749–752. doi: 10.1038/304749a0. [DOI] [PubMed] [Google Scholar]
- Sharp P. M., Li W. H. Ubiquitin genes as a paradigm of concerted evolution of tandem repeats. J Mol Evol. 1987;25(1):58–64. doi: 10.1007/BF02100041. [DOI] [PubMed] [Google Scholar]
- Siegelman M., Bond M. W., Gallatin W. M., St John T., Smith H. T., Fried V. A., Weissman I. L. Cell surface molecule associated with lymphocyte homing is a ubiquitinated branched-chain glycoprotein. Science. 1986 Feb 21;231(4740):823–829. doi: 10.1126/science.3003913. [DOI] [PubMed] [Google Scholar]
- Snutch T. P., Baillie D. L. Alterations in the pattern of gene expression following heat shock in the nematode Caenorhabditis elegans. Can J Biochem Cell Biol. 1983 Jun;61(6):480–487. doi: 10.1139/o83-064. [DOI] [PubMed] [Google Scholar]
- Sorger P. K., Lewis M. J., Pelham H. R. Heat shock factor is regulated differently in yeast and HeLa cells. Nature. 1987 Sep 3;329(6134):81–84. doi: 10.1038/329081a0. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Stone E. M., Rothblum K. N., Schwartz R. J. Intron-dependent evolution of chicken glyceraldehyde phosphate dehydrogenase gene. Nature. 1985 Feb 7;313(6002):498–500. doi: 10.1038/313498a0. [DOI] [PubMed] [Google Scholar]
- Struhl K. Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8419–8423. doi: 10.1073/pnas.82.24.8419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sulston J. E., Brenner S. The DNA of Caenorhabditis elegans. Genetics. 1974 May;77(1):95–104. doi: 10.1093/genetics/77.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urano Y., Watanabe K., Sakai M., Tamaoki T. The human albumin gene. Characterization of the 5' and 3' flanking regions and the polymorphic gene transcripts. J Biol Chem. 1986 Mar 5;261(7):3244–3251. [PubMed] [Google Scholar]
- Vijay-Kumar S., Bugg C. E., Cook W. J. Structure of ubiquitin refined at 1.8 A resolution. J Mol Biol. 1987 Apr 5;194(3):531–544. doi: 10.1016/0022-2836(87)90679-6. [DOI] [PubMed] [Google Scholar]
- Wiborg O., Pedersen M. S., Wind A., Berglund L. E., Marcker K. A., Vuust J. The human ubiquitin multigene family: some genes contain multiple directly repeated ubiquitin coding sequences. EMBO J. 1985 Mar;4(3):755–759. doi: 10.1002/j.1460-2075.1985.tb03693.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilkinson K. D., Cox M. J., O'Connor L. B., Shapira R. Structure and activities of a variant ubiquitin sequence from bakers' yeast. Biochemistry. 1986 Sep 9;25(18):4999–5004. doi: 10.1021/bi00366a005. [DOI] [PubMed] [Google Scholar]
- Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]