Analysis of the transcriptome of the protozoan Theileria parva using MPSS reveals that the majority of genes are transcriptionally active in the schizont stage.
Massively parallel signature sequencing (MPSS) was used to analyze the transcriptome of the intracellular protozoan Theileria parva. In total 1,095,000, 20 bp sequences representing 4371 different signatures were generated from T.parva schizonts. Reproducible signatures were identified within 73% of potentially detectable predicted genes and 83% had signatures in at least one MPSS cycle. A predicted leader peptide was detected on 405 expressed genes. The quantitative range of signatures was 4-52,256 transcripts per million (t.p.m.). Rare transcripts (<50 t.p.m.) were detected from 36% of genes. Sequence signatures approximated a lognormal distribution, as in microarray. Transcripts were widely distributed throughout the genome, although only 47% of 138 telomere-associated open reading frames exhibited signatures. Antisense signatures comprised 13.8% of the total, comparable with Plasmodium. Eighty five predicted genes with antisense signatures lacked a sense signature. Antisense transcripts were independently amplified from schizont cDNA and verified by sequencing. The MPSS transcripts per million for seven genes encoding schizont antigens recognized by bovine CD8 T cells varied 1000-fold. There was concordance between transcription and protein expression for heat shock proteins that were very highly expressed according to MPSS and proteomics. The data suggests a low level of baseline transcription from the majority of protein-coding genes.
Bishop
Richard
R
The International Livestock Research Institute (ILRI), PO Box 30709, Nairobi, Kenya. r.bishop@cgiar.org
Shah
Trushar
T
Pelle
Roger
R
Hoyle
David
D
Pearson
Terry
T
Haines
Lee
L
Brass
Andrew
A
Hulme
Helen
H
Graham
Simon P
SP
Taracha
Evans L N
EL
Kanga
Simon
S
Lu
Charles
C
Hass
Brian
B
Wortman
Jennifer
J
White
Owen
O
Gardner
Malcolm J
MJ
Nene
Vishvanath
V
de Villiers
Etienne P
EP
D000818:Animals; D018503:Genome, Protozoan; D023281:Genomics / Q000379:methods*; D016366:Open Reading Frames; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics; D016372:RNA, Antisense / Q000096:biosynthesis / Q000737:chemistry; D016053:RNA, Protozoan / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D017423:Sequence Analysis, RNA; D016615:Telomere / Q000737:chemistry; D016797:Theileria parva / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015533:Transcriptional Activation
D015800:Protozoan Proteins; D016372:RNA, Antisense; D016053:RNA, Protozoan