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A new twist in trypanosome RNA metabolism: cis-splicing of pre-mRNA

Published online by Cambridge University Press:  01 February 2000

GUNNAR MAIR
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
HUAFANG SHI
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
HONGJIE LI
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
APPOLINAIRE DJIKENG
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
HERNAN O. AVILES
Affiliation:
Indiana State University, Life Sciences Department, Terre-Haute, Indiana 47809, USA
JOSEPH R. BISHOP
Affiliation:
Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA
FRANCO H. FALCONE
Affiliation:
Institute of Cell, Animal and Population Biology, University of Edinburgh, Edinburgh EH9 3JT, United Kingdom
CRISTINA GAVRILESCU
Affiliation:
Department of Microbiology and Immunology, Cornell University, Ithaca, New York 14850, USA
JACQUI L. MONTGOMERY
Affiliation:
The Walter and Eliza Hall Institute, Post Office, Royal Melbourne Hospital, North Melbourne 3050, Australia
M. ISABEL SANTORI
Affiliation:
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina
LEAH S. STERN
Affiliation:
113B Veterans Administration Medical Center, San Francisco, California 94121, USA
ZEFENG WANG
Affiliation:
Department of Biological Chemistry, Johns Hopkins University, Baltimore, Maryland 21205, USA
ELISABETTA ULLU
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
CHRISTIAN TSCHUDI
Affiliation:
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8022, USA
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Abstract

It has been known for almost a decade and a half that in trypanosomes all mRNAs are trans-spliced by addition to the 5′ end of the spliced leader (SL) sequence. During the same time period the conviction developed that classical cis-splicing introns are not present in the trypanosome genome and that the trypanosome gene arrangement is highly compact with small intergenic regions separating one gene from the next. We have now discovered that these tenets are no longer true. Poly(A) polymerase (PAP) genes in Trypanosoma brucei and Trypanosoma cruzi are split by intervening sequences of 653 and 302 nt, respectively. The intervening sequences occur at identical positions in both organisms and obey the GT/AG rule of cis-splicing introns. PAP mRNAs are trans-spliced at the very 5′ end as well as internally at the 3′ splice site of the intervening sequence. Interestingly, 11 nucleotide positions past the actual 5′ splice site are conserved between the T. brucei and T. cruzi introns. Point mutations in these conserved positions, as well as in the AG dinucleotide of the 3′ splice site, abolish intron removal in vivo. Our results, together with the recent discovery of cis-splicing introns in Euglena gracilis, suggest that both trans- and cis-splicing are ancient acquisitions of the eukaryotic cell.

Type
Research Article
Information
RNA , Volume 6 , Issue 2 , February 2000 , pp. 163 - 169
Copyright
2000 RNA Society

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