Leishmania chagasi: the alpha-tubulin intercoding region results in constant levels of mRNA abundance despite protein synthesis inhibition and growth state.
The intercoding regions of many Leishmania sp. genes have been implicated in the regulation of mRNA processing, stability, and translation. Herein we show that the intercoding region of the Leishmania chagasi alpha-tubulin gene (alpha-TUB) confers stable beta-galactosidase (beta-GAL) reporter mRNA levels during promastigote growth and development in vitro and during protein synthesis inhibition. The abundance of both endogenous alpha-TUB mRNA and beta-GAL mRNA from a beta-GAL coding region situated upstream of the alpha-TUB intercoding region did not change significantly as promastigotes grew from logarithmic to stationary phase in vitro and the half-life of the beta-GAL mRNA remained constant. The abundance of both the endogenous alpha-TUB and the beta-GAL mRNA increased by less than 2-fold after protein synthesis inhibition corresponding to a moderate increase in mRNA half-life. These data suggest that the alpha-TUB intercoding region is an excellent control for the study of the regulation of other differentially expressed genes.
Purdy
Jay E
JE
Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA.
Donelson
John E
JE
Wilson
Mary E
ME
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D016054:DNA, Protozoan / Q000032:analysis / Q000302:isolation & purification; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D017930:Genes, Reporter; D006207:Half-Life; D006801:Humans; D018314:Leishmania infantum / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D008647:Mesocricetus; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D016053:RNA, Protozoan / Q000378:metabolism; D012692:Serial Passage; D014404:Tubulin / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D016054:DNA, Protozoan; D011500:Protein Synthesis Inhibitors; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D014404:Tubulin; D003513:Cycloheximide; D001616:beta-Galactosidase