Transcription of Leishmania major Friedlin chromosome 1 initiates in both directions within a single region.

Almost nothing is known about the sequences involved in transcription initiation of protein-coding genes in the parasite Leishmania. We describe here the transcriptional analysis of chromosome 1 (chr1) from Leishmania major Friedlin (LmjF) which encodes the first 29 genes on one DNA strand, and the remaining 50 on the opposite strand. Strand-specific nuclear run-on assays showed that a low level of nonspecific transcription probably takes place over the entire chromosome, but an approximately 10-fold higher level of coding strand-specific RNA polymerase II (Pol II)-mediated transcription initiates within the strand-switch region. 5' RACE studies localized the initiation sites to a <100 bp region. Transfection studies support the presence of a bidirectional promoter within the strand-switch region, but suggest that other factors are also involved in Pol II transcription. Thus, while in most eukaryotes each gene possesses its own promoter, a single region seems to drive the expression of the entire chr1 in LmjF.

[1]  L. Johnson,et al.  Transcriptional control elements of the rat thymidylate synthase promoter: evolutionary conservation of regulatory features. , 2000, Experimental cell research.

[2]  P. Borst,et al.  Alpha-amanitin-resistant transcription units in trypanosomes: a comparison of promoter sequences for a VSG gene expression site and for the ribosomal RNA genes. , 1991, Nucleic acids research.

[3]  R. Young,et al.  Transcription of eukaryotic protein-coding genes. , 2000, Annual review of genetics.

[4]  W. Sauerbier,et al.  Gene and transcription unit mapping by radiation effects. , 1978, Annual review of genetics.

[5]  S. Gallati,et al.  Overlapping sense and antisense transcription units in Trypanosoma brucei , 2001, Molecular microbiology.

[6]  D. Campbell,et al.  A conserved upstream element is essential for transcription of the Leishmania tarentolae mini‐exon gene. , 1994, The EMBO journal.

[7]  E. Pays,et al.  The actin gene promoter of Trypanosoma brucei. , 1991, Nucleic acids research.

[8]  P. Myler,et al.  The unusual gene organization of Leishmania major chromosome 1 may reflect novel transcription processes. , 2000, Nucleic acids research.

[9]  P. Myler,et al.  Sense and antisense transcripts in the histone H1 (HIS-1) locus of Leishmania major. , 2003, International journal for parasitology.

[10]  P. Borst,et al.  The anatomy and transcription of a telomeric expression site for variant-specific surface antigens in T. brucei , 1987, Cell.

[11]  J. Donelson,et al.  Comparison of the Post-transcriptional Regulation of the mRNAs for the Surface Proteins PSA (GP46) and MSP (GP63) ofLeishmania chagasi * , 2002, The Journal of Biological Chemistry.

[12]  S. Sunkin,et al.  Genomic organization and functional characterization of the Leishmania major Friedlin ribosomal RNA gene locus. , 2001, Molecular and biochemical parasitology.

[13]  M. Fried,et al.  The bidirectional promoter of the divergently transcribed mouse Surf-1 and Surf-2 genes. , 1991, Molecular and cellular biology.

[14]  J. Ellis,et al.  The Leishmania major RNA polymerase II largest subunit lacks a carboxy-terminus heptad repeat structure and its encoding gene is linked with the calreticulin gene. , 2000, Protist.

[15]  S. Sunkin,et al.  Leishmania major Friedlin chromosome 1 has an unusual distribution of protein-coding genes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  E. Pays,et al.  Ultraviolet irradiation inhibits RNA decay and modifies ribosomal RNA processing in Trypanosoma brucei. , 1991, Molecular and biochemical parasitology.

[17]  David Hirsh,et al.  A trans-spliced leader sequence on actin mRNA in C. elegans , 1987, Cell.

[18]  D. Salmon,et al.  The genes and transcripts of an antigen gene expression site from T. brucei , 1989, Cell.

[19]  P. Bastien,et al.  The switch region on Leishmania major chromosome 1 is not required for mitotic stability or gene expression, but appears to be essential. , 2002, Nucleic acids research.

[20]  B. Barrell,et al.  Secondary DNA structure analysis of the coding strand switch regions of five Leishmania major Friedlin chromosomes , 2001, Current Genetics.

[21]  S. Beverley,et al.  Nuclease mapping and DNA sequence analysis of transcripts from the dihydrofolate reductase-thymidylate synthase (R) region of Leishmania major. , 1990, Nucleic acids research.

[22]  P. Borst,et al.  Post-transcriptional control of the differential expression of phosphoglycerate kinase genes in Trypanosoma brucei. , 1988, Journal of molecular biology.

[23]  S. Sunkin,et al.  Genomic organization and gene function in Leishmania. , 2000, Biochemical Society transactions.

[24]  J. Donelson,et al.  Search for promoters for the GARP and rRNA genes of Trypanosoma congolense. , 1999, Molecular and biochemical parasitology.

[25]  D. Wirth,et al.  Identification of a cis-acting gene regulatory element from the lemdr1 locus of Leishmania enriettii. , 1994, The Journal of biological chemistry.

[26]  C. Clayton,et al.  Life without transcriptional control? From fly to man and back again , 2002, The EMBO journal.

[27]  J. Barry,et al.  A promotor directing alpha-amanitin-sensitive transcription of GARP, the major surface antigen of insect stage Trypanosoma congolense. , 1996, Nucleic acids research.

[28]  C. Clayton,et al.  Testing promoter activity in the trypanosome genome: isolation of a metacyclic-type VSG promoter, and unexpected insights into RNA polymerase II transcription. , 1998, Experimental parasitology.

[29]  N. Agabian Trans splicing of nuclear pre-mRNAs , 1990, Cell.

[30]  L. V. D. van der Ploeg,et al.  RNA polymerase I-mediated protein-coding gene expression in Trypanosoma brucei. , 1992, Parasitology today.

[31]  S. Sunkin,et al.  A low-background inducible promoter system in Leishmania donovani. , 2002, Molecular and biochemical parasitology.

[32]  Jean Thierry-Mieg,et al.  A global analysis of Caenorhabditis elegans operons , 2002, Nature.

[33]  Yang Li,et al.  A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA polymerase II , 1994, Cell.

[34]  P. Borst,et al.  Inactivation of transcription by UV irradiation of T. brucei provides evidence for a multicistronic transcription unit including a VSG gene , 1987, Cell.

[35]  V. Bellofatto,et al.  Trypanosome spliced leader RNA genes contain the first identified RNA polymerase II gene promoter in these organisms. , 2001, Nucleic acids research.

[36]  L. Johnson,et al.  Transcriptional control elements and complex initiation pattern of the TATA‐less bidirectional human thymidylate synthase promoter , 2000, Journal of cellular biochemistry.

[37]  M. Lee An RNA polymerase II promoter in the hsp70 locus of Trypanosoma brucei , 1996, Molecular and cellular biology.

[38]  M. Parsons,et al.  Molecular biology of parasitic protozoa , 1996 .