Cotranscription and intergenic splicing of human galactose-1-phosphate uridylyltransferase and interleukin-11 receptor alpha-chain genes generate a fusion mRNA in normal cells. Implication for the production of multidomain proteins during evolution.

In the past 10 years, much attention has been focused on transcription preinitiation complex formation as a target for regulating gene expression, and other targets such as transcription termination complex assemblage have been less intensively investigated. We established the existence of poly(A) site choice and fusion splicing of two adjacent genes, galactose-1-phosphate uridylyltransferase (GALT) and interleukin-11 receptor alpha-chain (IL-11Ralpha), in normal human cells. This 16-kilobase (kb) transcription unit contains two promoters (the first one is constitutive, and the second one, 8 kb downstream, is highly regulated) and two cleavage/polyadenylation signals separated by 12 kb. The promoter from the GALT gene yields two mRNAs, a 1.4-kb mRNA encoding GALT and a 3-kb fusion mRNA when the first poly(A) site is spliced out and the second poly(A) is used. The 3-kb mRNA codes for a fusion protein of unknown function, containing part of the GALT protein and the entire IL-11Ralpha protein. The GALT promoter/IL-11Ralpha poly(A) transcript results from leaky termination and alternative splicing. This feature of RNA polymerase (pol) II transcription, which contrasts with efficient RNA pol I and pol III termination, may be involved, together with chromosome rearrangements, in the generation of fusion proteins with multiple domains and would have major evolutionary implications in terms of natural processes to generate novel proteins with common motifs. Our results, together with accumulation of genomic informations, will stimulate new considerations and experiments in gene expression studies.

[1]  A. Hirano,et al.  Structure and function of bicistronic RNA encoding the phosphoprotein and matrix protein of measles virus , 1987, Journal of virology.

[2]  B. Seed,et al.  Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselection procedure. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Kolakofsky,et al.  Transcriptional map of the measles virus genome. , 1986, The Journal of general virology.

[4]  Charles R.scriver,et al.  The Metabolic basis of inherited disease , 1989 .

[5]  C. Begley,et al.  Cloning of a murine IL‐11 receptor alpha‐chain; requirement for gp130 for high affinity binding and signal transduction. , 1994, The EMBO journal.

[6]  Tedesco Ta,et al.  An improved assay of erythrocyte and leukocyte galactose-1-phosphate uridyl transferase: stabilization of the enzyme by a thiol protective reagent. , 1965 .

[7]  K. Arai,et al.  Structure of the gene encoding the alpha subunit of the human granulocyte-macrophage colony stimulating factor receptor. Implications for the evolution of the cytokine receptor superfamily. , 1994, The Journal of biological chemistry.

[8]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[9]  W. Wold,et al.  Genetic analysis of mRNA synthesis in adenovirus region E3 at different stages of productive infection by RNA-processing mutants , 1986, Journal of virology.

[10]  R. Gibbs,et al.  Large-scale sequencing in human chromosome 12p13: experimental and computational gene structure determination. , 1997, Genome research.

[11]  M. Bonneville,et al.  HLA-target antigens and T-cell receptor diversity of activated T cells invading the skin during acute graft-versus-host disease. , 1996, Blood.

[12]  M. Steinmetz,et al.  IL3-dependent mouse clones that express B-220 surface antigen, contain ig genes in germ-line configuration, and generate B lymphocytes in vivo , 1985, Cell.

[13]  G. Peters,et al.  Complete nucleotide sequence of a milk-transmitted mouse mammary tumor virus: two frameshift suppression events are required for translation of gag and pol , 1987, Journal of virology.

[14]  J. Rowley,et al.  Intergenic splicing of MDS1 and EVI1 occurs in normal tissues as well as in myeloid leukemia and produces a new member of the PR domain family. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  N. Leslie,et al.  The human galactose-1-phosphate uridyltransferase gene. , 1992, Genomics.

[16]  W. Alexander,et al.  Twenty proteins containing a C-terminal SOCS box form five structural classes. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Heinrich,et al.  Reconstitution of two isoforms of the human interleukin‐11 receptor and comparison of their functional properties , 1997 .

[18]  E. Bateman,et al.  Promoter occlusion during ribosomal RNA transcription , 1988, Cell.

[19]  M. Ultsch,et al.  Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. , 1992, Science.

[20]  D. Williams,et al.  Interleukin-11: review of molecular, cell biology, and clinical use. , 1997, Blood.

[21]  G. Elliott,et al.  Virus and host cell-dependent variation in transcription of the mumps virus genome. , 1990, The Journal of general virology.

[22]  B. Cullen,et al.  Transcriptional interference in avian retroviruses—implications for the promoter insertion model of leukaemogenesis , 1984, Nature.

[23]  P. Pearson,et al.  Assignment of a human galactose-1-phosphate uridylyltransferase gene (GALT1) to chromosome 9 in human-Chinese hamster somatic cell hybrids. , 1978, Cytogenetics and cell genetics.

[24]  N. Proudfoot.  Transcriptional interference and termination between duplicated α-globin gene constructs suggests a novel mechanism for gene regulation , 1986, Nature.

[25]  J. Concordet,et al.  Illegitimate transcription: transcription of any gene in any cell type. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Y. Yang,et al.  Interleukin-11 and its receptor. , 1992, BioFactors.

[27]  C J Rawlings,et al.  Computational gene discovery and human disease. , 1997, Current opinion in genetics & development.

[28]  G. Wertz,et al.  The Product of the Respiratory Syncytial Virus M2 Gene ORF1 Enhances Readthrough of Intergenic Junctions during Viral Transcription , 1998, Journal of Virology.

[29]  R. Bataille,et al.  Molecular cloning of two isoforms of a receptor for the human hematopoietic cytokine interleukin-11. , 1995, Blood.

[30]  D. Spector,et al.  Readthrough activation of early adenovirus E1b gene transcription , 1997, Journal of virology.

[31]  J. Coffin,et al.  Influence of sequences in the long terminal repeat and flanking cell DNA on polyadenylation of retroviral transcripts , 1993, Journal of virology.

[32]  C. Begley,et al.  The human IL-11 receptor requires gp130 for signalling: demonstration by molecular cloning of the receptor. , 1996, Oncogene.

[33]  B. Falk,et al.  The beet western yellows virus ST9-associated RNA shares structural and nucleotide sequence homology with carmo-like viruses. , 1993, Virology.

[34]  T. Morrison,et al.  Structural and functional characterization of Newcastle disease virus polycistronic RNA species , 1984, Journal of virology.

[35]  F. van Leuven,et al.  Molecular cloning and characterization of the human interleukin-11 receptor alpha-chain gene, IL11RA, located on chromosome 9p13. , 1996, Genomics.

[36]  P. Frey,et al.  Three-dimensional structure of galactose-1-phosphate uridylyltransferase from Escherichia coli at 1.8 A resolution. , 1995, Biochemistry.

[37]  J. Bazan,et al.  Structural design and molecular evolution of a cytokine receptor superfamily. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[38]  I. Peake.  The Molecular Basis of Blood Diseases , 1988 .

[39]  S. Martin,et al.  Characterization of clones for the sixth (L) gene and a transcriptional map for morbilliviruses. , 1986, The Journal of general virology.

[40]  K. Murti,et al.  Elevated expression of the human parainfluenza virus type 1 F gene downregulates HN expression. , 1997, Virology.

[41]  H. Kalckar.  50 years of biological research--from oxidative phosphorylation to energy requiring transport regulation. , 1991, Annual review of biochemistry.

[42]  J. Mallet,et al.  Specific Vesicular Acetylcholine Transporter Promoters Lie within the First Intron of the Rat Choline Acetyltransferase Gene (*) , 1995, The Journal of Biological Chemistry.

[43]  R. Kopito.  ER Quality Control: The Cytoplasmic Connection , 1997, Cell.

[44]  William McGinnis,et al.  Homeobox genes and axial patterning , 1992, Cell.

[45]  B. Quimby,et al.  Functional Requirements of the Active Site Position 185 in the Human Enzyme Galactose-1-phosphate Uridylyltransferase* , 1996, The Journal of Biological Chemistry.

[46]  G. Milligan,et al.  Antibodies directed against synthetic peptides distinguish between GTP-binding proteins in neutrophil and brain. , 1987, The Journal of biological chemistry.