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Inosine and N1-methylinosine within a synthetic oligomer mimicking the anticodon loop of human tRNAAla are major epitopes for anti-PL-12 myositis autoantibodies

Published online by Cambridge University Press:  01 July 1999

H.F. BECKER ,
Y. CORDA ,
M.B. MATHEWS ,
J.-L. FOURREY  and
H. GROSJEAN
H.F. BECKER
Affiliation:
Laboratoire d'Enzymologie et Biochimie Structurales du Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, F-91198, Gif-sur-Yvette, France Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, F-91198, Gif-sur-Yvette, France Present address: Université de Paris 6, Laboratoire de Chimie Organique Biologique, 4 place Jussieu, F-75252 Cedex 05, France.
Y. CORDA
Affiliation:
Laboratoire d'Enzymologie et Biochimie Structurales du Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, F-91198, Gif-sur-Yvette, France Present address: CNRS, Centre de Biologie Moléculaire, 31 chemin J. Aiguier, F-13402 Marseille Cedex 20, France.
M.B. MATHEWS
Affiliation:
Department of Biochemistry and Molecular Biology, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103-2714, USA
J.-L. FOURREY
Affiliation:
Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, F-91198, Gif-sur-Yvette, France
H. GROSJEAN
Affiliation:
Laboratoire d'Enzymologie et Biochimie Structurales du Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, F-91198, Gif-sur-Yvette, France
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Abstract

Sera of some patients afflicted with the inflammatory disease myositis contain antibodies of the anti-PL-12 type. A fraction of these polyclonal autoantibodies specifically precipitates the fully matured human tRNAAla bearing the anticodon IGC (PL-12 antigen). Earlier work (Bunn & Mathews, 1987, Science 238:116–119) had shown that the epitopes are located entirely within the anticodon stem-loop of the tRNAAla. Here we demonstrate that human anti-tRNAAla autoantibodies immunoprecipitate a synthetic polyribonucleotide containing inosine (I) and N1-methylinosine (m1I) separated by 2 nt as in the anticodon stem-loop of human tRNAAla. The shortest polyribonucleotide that can be immunoprecipitated corresponds to the pentanucleotide IpGpCpm1IpUp, which corresponds to part of the anticodon loop of human tRNAAla and lacks the stem-loop structure. The efficiency of immunoprecipitation was about four times greater with longer polyribonucleotides capable of forming a stem-loop structure, and was abolished by altering the relative positions of I and m1I within the synthetic polynucleotide. Synthetic oligodeoxyribonucleotide analogs of the tRNAAla stem-loop, containing the sequence dIpdGdCdm1Ip, are not antigenic. Our results show that human anti-tRNAAla autoantibodies selectively recognize chemical details of modified nucleotides (the 6-keto group of inosine-34 and the 6-keto group and the N1-methyl groups of N1-methylinosine-37) within an anticodon loop structure of a tRNA molecule. We also describe the chemical synthesis of the phosphoramidite derivatives corresponding to N1-methylinosine and N1-methyl-2′-deoxyinosine for use in the automatic chemical synthesis of oligonucleotides containing N1-methylinosine and N1-methyl-2′-deoxyinosine.

Keywords

antigenautoantibodiesepitopeimmunoprecipitationinosinemyositisN1-methylinosinephosphoramiditetRNA
Type
Research Article
Information
RNA , Volume 5 , Issue 7 , July 1999 , pp. 865 - 875
Copyright
© 1999 RNA Society

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Footnotes

The results of this paper were first reported at the Third Dresden Symposium on Autoantibodies held in Dresden on September 26\N28, 1996.