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Eukaryotic Cell, October 2003, p. 1009-1017, Vol. 2, No. 5
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.5.1009-1017.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Negative Regulatory Element Controls mRNA Abundance of the Leishmania mexicana Paraflagellar Rod Gene PFR2{dagger}

Krishna K. Mishra, Timothy R. Holzer, Landon L. Moore,{ddagger} and Jonathan H. LeBowitz*

Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907-2063

Received 14 January 2003/ Accepted 7 July 2003

The Leishmania mexicana PFR2 locus encodes a component of the paraflagellar rod (PFR), a flagellar structure found only in the insect stage of the life cycle. PFR2 mRNA levels are 10-fold lower in the mammalian stage than in the insect stage. Nuclear run-on experiments indicate that the change in PFR2 mRNA abundance is achieved posttranscriptionally. Deletion and block substitution analysis of the entire 1,400-nucleotide 3' untranslated region (UTR) of PFR2C led to the identification of a regulatory element contained within 10 nucleotides of the 3' UTR, termed the PFR regulatory element (PRE), that is necessary for the 10-fold regulation of PFR2 mRNA levels. Comparison of the half-lives of PFR2 transcripts, identical except for the presence or absence of the PRE, revealed that the PRE acts by destabilizing the PFR2 mRNA in amastigotes. The PRE was inserted into a construct which directs the constitutive expression of a chimeric PFR2 transcript. Insertion of the PRE resulted in regulated expression of this transcript, demonstrating that the regulatory element is sufficient for promastigote-specific expression. Since the PRE is present in the 3' UTR of all L. mexicana PFR genes examined so far, we propose that it serves a means of coordinating expression of PFR genes.

* Corresponding author. Present address: Symbiontics, Inc., 4041 Forest Park Ave., St. Louis, MO 63108. Phone: (314) 615-6966. Fax: (314) 615-6901. E-mail: jon{at}emergingtech.org.

{dagger} This is paper 16976 from Purdue Agriculture Research Programs.

{ddagger} Present address: Department of Genetics and Genomics, Boston University School of Medicine, Boston, MA 02118.

Eukaryotic Cell, October 2003, p. 1009-1017, Vol. 2, No. 5
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.5.1009-1017.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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