Rapid decay of unstable Leishmania mRNAs bearing a conserved retroposon signature 3'-UTR motif is initiated by a site-specific endonucleolytic cleavage without prior deadenylation.
We have previously shown that the Leishmania genome possess two widespread families of extinct retroposons termed Short Interspersed DEgenerated Retroposons (SIDER1/2) that play a role in post-transcriptional regulation. Moreover, we have demonstrated that SIDER2 retroposons promote mRNA degradation. Here we provide new insights into the mechanism by which unstable Leishmania mRNAs harboring a SIDER2 retroposon in their 3'-untranslated region are degraded. We show that, unlike most eukaryotic transcripts, SIDER2-bearing mRNAs do not undergo poly(A) tail shortening prior to rapid turnover, but instead, they are targeted for degradation by a site-specific endonucleolytic cleavage. The main cleavage site was mapped in two randomly selected SIDER2-containing mRNAs in vivo between an AU dinucleotide at the 5'-end of the second 79-nt signature (signature II), which represents the most conserved sequence amongst SIDER2 retroposons. Deletion of signature II abolished endonucleolytic cleavage and deadenylation-independent decay and increased mRNA stability. Interestingly, we show that overexpression of SIDER2 anti-sense RNA can increase sense transcript abundance and stability, and that complementarity to the cleavage region is required for protecting SIDER2-containing transcripts from degradation. These results establish a new paradigm for how unstable mRNAs are degraded in Leishmania and could serve as the basis for a better understanding of mRNA decay pathways in general.
Müller
Michaela
M
Infectious Disease Research Centre, CHUL Research Centre and Department of Microbiology and Immunology, Faculty of Medicine, Laval University, Quebec, Canada.
Padmanabhan
Prasad K
PK
Rochette
Annie
A
Mukherjee
Debdutta
D
Smith
Martin
M
Dumas
Carole
C
Papadopoulou
Barbara
B
D020413:3' Untranslated Regions; D001483:Base Sequence; D017124:Conserved Sequence; D004722:Endoribonucleases / Q000378:metabolism*; D018320:Leishmania major / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D020871:RNA Stability; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D018626:Retroelements
D020413:3' Untranslated Regions; D016372:RNA, Antisense; D012333:RNA, Messenger; D018626:Retroelements; D004722:Endoribonucleases
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