Journal article
Regulation of genes encoding the major surface protease of Leishmania chagasi via mRNA stability
Molecular and biochemical parasitology, Vol.142(1), pp.88-97
07/2005
DOI: 10.1016/j.molbiopara.2005.03.010
PMID: 15876463
Abstract
The intercoding regions between many Leishmania sp. genes regulate their mRNA expression. The MSPL mRNA, encoding a subclass of the major surface protease (MSP) of Leishmania chagasi, increases in abundance, when protein synthesis is arrested, while alpha-tubulin (alpha-TUB) mRNA and most other mRNAs do not. We found that the intercoding region between MSPL-coding regions, when cloned downstream of the beta-galactosidase reporter gene (beta-GAL), caused beta-GAL mRNA to increase 8- to 10-fold after inhibiting protein synthesis with cycloheximide. Stable L. chagasi transfectants containing hybrid MSPL/alpha-TUB intercoding regions cloned downstream of beta-GAL were made. The alpha-TUB intercoding region induced high-level baseline beta-GAL mRNA that increased only 1.3-fold after incubation with cycloheximide. In contrast, the MSPL intercoding region, as well as constructs containing nucleotides 303-505 from the MSPL 3'UTR, caused steady-state beta-GAL mRNA levels in the absence of cycloheximide that were approximately 10% of alpha-TUB constructs. These levels increased between 4.4- and 13.2-fold after cycloheximide was added. Constructs containing half of this region (303-394 or 395-505) produced intermediate levels of beta-GAL mRNA and intermediate levels of cycloheximide induction. The kinetics of cycloheximide induction of beta-GAL mRNA was similar with region 303-505 constructs as with constructs bearing the entire endogenous MSPL intercoding region. Furthermore, region 303-505 increased reporter mRNA abundance after cycloheximide by increasing mRNA half-life. Hence, we have identified a 202-nucleotide region within the MSPL 3'UTR that is in part responsible for cycloheximide induction. We hypothesize that this region may interact with labile regulatory protein factor(s).
Details
- Title: Subtitle
- Regulation of genes encoding the major surface protease of Leishmania chagasi via mRNA stability
- Creators
- Jay E Purdy - Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA. jay-purdy@uiowa.eduJohn E DonelsonMary E Wilson
- Resource Type
- Journal article
- Publication Details
- Molecular and biochemical parasitology, Vol.142(1), pp.88-97
- Publisher
- Netherlands
- DOI
- 10.1016/j.molbiopara.2005.03.010
- PMID
- 15876463
- ISSN
- 0166-6851
- eISSN
- 1872-9428
- Grant note
- AI07343 / NIAID NIH HHS R01 AI45540 / NIAID NIH HHS R01 AI32135 / NIAID NIH HHS K08 AI055804-01 / NIAID NIH HHS R01 AI48822 / NIAID NIH HHS
- Language
- English
- Date published
- 07/2005
- Academic Unit
- Microbiology and Immunology; International Programs; Epidemiology; Biochemistry and Molecular Biology; Internal Medicine
- Record Identifier
- 9984001220802771
Metrics
16 Record Views