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PMID: 2243782 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Nuclease mapping and DNA sequence analysis of transcripts from the dihydrofolate reductase-thymidylate synthase (R) region of Leishmania major.

Nucleic acids research ·Vol. 18 ·No. 21 ·1990-11-11 ·Pages 6399-408

Kapler GM, Zhang K, Beverley SM

Abstract

Trypanosomatid protozoan parasites utilize a number of nonstandard mechanisms in expressing their genes. To probe these phenomena in a genetically accessible system, we have mapped termini of eight transcripts arising from the amplified R region including the DHFR-TS gene of methotrexate-resistant Leishmania major. Poly(A)+ RNAs transcribed from the DHFR-TS-coding strand exhibit features similar to those observed around other trypanosomatid protein-coding genes. These include close spacing, the presence of a transpliced miniexon on the 5' termini, heterogeneity at both 5' and 3' ends, and in some cases S1 nuclease protection of intertranscript regions. Other than the splice acceptor site, no consensus sequence elements associated with either 5' or 3' ends were detected, although polydinucleotide tracts tended to be near inter-transcript regions. Two poly(A)+ RNAs transcribed from the opposite strand of the upstream flanking regions lacked the miniexon. Sequencing of DNA encoding the overlapping 1.7 kb opposite strand transcripts (one bearing and one lacking the miniexon, both found on polysomes) revealed no reading frames likely to encode proteins, suggesting that at least some of these RNAs could be nonfunctional by-products of RNA processing.

MeSH Terms
Animals Base Sequence DNA DNA, Protozoan/genetics Genes Leishmania tropica/enzymology,genetics Molecular Sequence Data Nucleotide Mapping Oligonucleotide Probes Polymerase Chain Reaction/methods RNA Splicing RNA, Protozoan/genetics,isolation & purification Restriction Mapping Sequence Homology, Nucleic Acid Single-Strand Specific DNA and RNA Endonucleases Tetrahydrofolate Dehydrogenase/genetics Thymidylate Synthase/genetics Transcription, Genetic
Chemicals
DNA, Protozoan Oligonucleotide Probes RNA, Protozoan DNA Tetrahydrofolate Dehydrogenase Thymidylate Synthase Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kapler G M
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Zhang K
Beverley S M
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.21.6399
Published
1990-11-11
Pages
6399-408
Language
English
Region
England
NLM ID
0411011
PMCID
PMC332520
Subset
IM
Grants
NIGMS NIH HHS · 5T-32-GM07306 · United States
NIAID NIH HHS · AI-2903 · United States
NCRR NIH HHS · S07 RR 05381-27 · United States
Databases
GENBANK
X51733, X51734
External Links
PubMed source DOI: 10.1093/nar/18.21.6399 PMC Full Text View journal details
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