Distinct 3′-Untranslated Region Elements Regulate Stage-specific mRNA Accumulation and Translation in Leishmania

We recently characterized a large developmentally regulated gene family in Leishmania encoding the amastin surface proteins. While studying the regulation of these genes, we identified a region of 770 nucleotides (nt) within the 2055-nt 3′-untranslated region (3′-UTR) that regulates stage-specific gene expression at the level of translation. An intriguing feature of this 3′-UTR regulatory region is the presence of a ∼450-nt element that is highly conserved among several Leishmania mRNAs. Here we show, using a luciferase reporter system that the element of the amastin in is the that the the a region of nucleotides at the of several amastin in 3′-UTR regulatory in amastin translation. we that in the of the of the of amastin a that is of the these the that stage-specific regulation of the amastin in Leishmania is the of that of the within We recently characterized a large developmentally regulated gene family in Leishmania encoding the amastin surface proteins. While studying the regulation of these genes, we identified a region of 770 nucleotides (nt) within the 2055-nt 3′-untranslated region (3′-UTR) that regulates stage-specific gene expression at the level of translation. An intriguing feature of this 3′-UTR regulatory region is the presence of a ∼450-nt element that is highly conserved among several Leishmania mRNAs. Here we show, using a luciferase reporter system that the element of the amastin in is the that the the a region of nucleotides at the of several amastin in 3′-UTR regulatory in amastin translation. we that in the of the of the of amastin a that is of the these the that stage-specific regulation of the amastin in Leishmania is the of that of the within Leishmania a in several the of Leishmania the of is Leishmania the is in several the of of of Leishmania the the the the within the of the a within the that the expression of a of several characterized in Leishmania in the of in stage-specific regulation of gene expression at the level in within the 3′-UTR of that the stage-specific regulation of gene expression in this that of in the of the the of in expression that of the Leishmania in the in in the of in gene expression in this identified a developmentally regulated gene family in Leishmania encoding the amastin surface that the amastin We recently characterized this large gene family in Leishmania that the of in the of the regulation of the amastin in Leishmania in is within the 3′-UTR that in in While studying of the amastin gene we a region of 770 within the 3′-UTR that regulates stage-specific gene expression at the level of An intriguing feature of this 3′-UTR regulatory region is the presence of a ∼450-nt element that is highly conserved among the of the amastin among several Leishmania we that the element of the amastin in is the that the the the we that a region of at the of several amastin amastin in we that in the the of of the amastin the of amastin in the a that is of the the that stage-specific regulation of the amastin in Leishmania is the of that within Leishmania in Leishmania in this at in in a the of in in at in these of in of of the Leishmania luciferase of the in of the the in of the 3′-UTR regulatory region of the amastin gene using the of of the luciferase a region of nucleotides the conserved element nucleotides using the element the nucleotides of the amastin 3′-UTR the nucleotides of the amastin 3′-UTR using the nucleotides of the region a the the of the amastin the the gene the region of the amastin in the in the in of that the of the gene the of of the of of of the amastin 3′-UTR regulatory in stage-specific of the the of 3′-UTR regulatory of the amastin of the region the region the is is the 3′-UTR regulatory the of the of the of the the a the the Leishmania is the in a the region the of 3′-UTR regulatory in of the expression of the amastin using a the amastin 3′-UTR of luciferase reporter the the luciferase the of the of using the Leishmania using of a in the in a a at the using a using a An a in Leishmania at at the of a in the in a at at at using of at of the using 3′-UTR of the recently characterized a large family of developmentally regulated in surface of the amastin gene that we in we identified a region of 770 in the 3′-UTR that regulates stage-specific gene expression at the level of the nucleotides of this regulatory region conserved among several in the of the amastin the of the in several of that of the element We that this conserved element the regulation of the amastin this we a of within the region that we of the luciferase reporter gene Leishmania of expression in in using in the region in in element in of the Leishmania the of the amastin gene that the region of a nucleotides of of the element this we a that of of the of the regulation the region the nucleotides that the element is regulation that the of the element that within the region of the element in regulation of the amastin this we the of the region the region this of the of in the the of regulation the element in of nucleotides of of the region in regulation the regulatory of the amastin region of in the region the element in the Leishmania conserved in Leishmania amastin gene is that a within the region regulation that this in several mRNAs. within this region this in of the of luciferase reporter we that within the amastin 3′-UTR region the of in the of the regulation in of these regulatory the of the 3′-UTR stage-specific we of the in in the the the in that of the 3′-UTR that the regulation these that the of 3′-UTR identified within the region of the amastin is in is the of of 3′-UTR amastin translation. of the in of a a the luciferase the of the using the that amastin in Leishmania in is that is the nucleotides of the amastin we the that the within the 770 nucleotides of the 3′-UTR a in this the of the the of the amastin that in we a of of the regulatory 3′-UTR gene the of Leishmania the gene the region the the region the element the the a Leishmania is of of in the the of the amastin in these we that the in amastin in the the the luciferase reporter the of the 3′-UTR the of a of these in the 3′-UTR highly we the of 3′-UTR regulatory of the amastin the element element the that a at the these a the of the amastin 3′-UTR regulatory that the the a a of of the in the the the is the at in a in the of the the of the that is the of the 3′-UTR regulatory is in in the in of the amastin 3′-UTR regulatory in of a the a at the the of the the of the in the of a the of the of in the of in the of the of the of in a a of the in a of the regulation of stage-specific gene expression in Leishmania is in the in that 3′-UTR a in the of in Leishmania is a of that the of the of of the of the amastin in we the of Leishmania the gene the amastin the 3′-UTR regulatory Leishmania the the in the of these the of amastin that of the amastin in the regulation of the amastin in the of the in the amastin the of the amastin 3′-UTR the the of the regulation of the amastin using a luciferase reporter the amastin the in the luciferase the of of a the the of Leishmania in the in the of is the of the we these regulation of amastin mRNAs. of the 3′-UTR regulatory in of the the of of the in within in the the at the of the these we that regulation is Leishmania the the of Leishmania within that amastin translation. regulation of the amastin the the the a of the of these the luciferase in a at at the of amastin a that is of the the the in the of the amastin the of of the that the of amastin in the Here we that 3′-UTR that amastin in in these of of the amastin that is a 2055-nt 3′-UTR in that stage-specific amastin gene expression of that within we a of regulation that 3′-UTR of the developmentally regulated amastin in We these of the within is gene regulation in this the that stage-specific regulation of gene expression in that regulation of the amastin is the 770 nucleotides of the a ∼450-nt element that is highly conserved among several Leishmania a of We that this 3′-UTR element in stage-specific Here we that the element of the amastin in the the that of the of the the this element is a of the the of is that the element that the of the the in this that regulation the conserved element the region of a of of of the that is that of the the of in of these the the presence of that conserved among the of the element the that several Leishmania regulated a of in of regulation a the a within is that the regulation is a the is the the of that stage-specific regulation in is that amastin is 3′-UTR in the a region of that is in of the amastin is in the this identified region in of the regulatory element the a of this in is that the of in that in of the amastin at regulation at the level of a the of the amastin surface the of the in the presence of a that of translation. the of the 3′-UTR in the the that regulation these at the level of is the of regulation in is the of in of 3′-UTR of several of in of the of the of a a at the of the 3′-UTR in identified the the in this is that regulation of the amastin gene family is at the level of at the level of that within is that the of regulation that the 3′-UTR regulatory of amastin mRNAs. system of we that of these 3′-UTR regulatory a in amastin regulation that the of amastin in the of the that this is in the of the in amastin is highly the the in the that translation. in that in of of that a of in in the in this a of regulation the developmentally regulated amastin gene family in Leishmania that is in within a regulation that is in a regulatory element within the is in is the that the of regulation that stage-specific gene expression in the of these gene regulation in of in the in a in within the 3′-UTR the in in in a in within the 3′-UTR the in in in the of in in within the 3′-UTR the in in in a in within the 3′-UTR the in in in in in a Leishmania a in several the of Leishmania the of is Leishmania the is in several the of of of Leishmania the the the the within the of the a within the that the expression of a of several characterized in Leishmania in the of in stage-specific regulation of gene expression at the level in within the 3′-UTR of that the stage-specific regulation of gene expression in this that of in the of the the of in expression that of the Leishmania in the in in the of in gene expression in this region region We identified a developmentally regulated gene family in Leishmania encoding the amastin surface that the amastin We recently characterized this large gene family in Leishmania that the of in the of the regulation of the amastin in Leishmania in is within the 3′-UTR that in in While studying of the amastin gene we a region of 770 within the 3′-UTR that regulates stage-specific gene expression at the level of An intriguing feature of this 3′-UTR regulatory region is the presence of a ∼450-nt element that is highly conserved among the of the amastin among several Leishmania we that the element of the amastin in is the that the the the we that a region of at the of several amastin amastin in we that in the the of of the amastin the of amastin in the a that is of the the that stage-specific regulation of the amastin in Leishmania is the of that within Leishmania in Leishmania in this at in in a the of in in at in these of in of of the Leishmania luciferase of the in of the the in of the 3′-UTR regulatory region of the amastin gene using the of of the luciferase a region of nucleotides the conserved element nucleotides using the element the nucleotides of the amastin 3′-UTR the nucleotides of the amastin 3′-UTR using the nucleotides of the region a the the of the amastin the the gene the region of the amastin in the in the in of that the of the gene the of of the of of the of using the Leishmania using of a in the in a a at the using a using a An a in Leishmania at at the of a in the in a at at at using of at of the using Leishmania in Leishmania in this at in in a the of in in at in these of in of of the Leishmania luciferase of the in of the the in of the 3′-UTR regulatory region of the amastin gene using the of of the luciferase a region of nucleotides the conserved element nucleotides using the element the nucleotides of the amastin 3′-UTR the nucleotides of the amastin 3′-UTR using the nucleotides of the region a the the of the amastin the the gene the region of the amastin in the in the in of that the of the gene the of of the of of the of using the Leishmania using of a in the in a a at the using a using a An a in Leishmania at at the of a in the in a at at at using of at of the using 3′-UTR of the recently characterized a large family of developmentally regulated in surface of the amastin gene that we in we identified a region of 770 in the 3′-UTR that regulates stage-specific gene expression at the level of the nucleotides of this regulatory region conserved among several in the of the amastin the of the in several of that of the element We that this conserved element the regulation of the amastin this we a of within the region that we of the luciferase reporter gene Leishmania of expression in in using in the region in in element in of the Leishmania the of the amastin gene that the region of a nucleotides of of the element this we a that of of the of the regulation the region the nucleotides that the element is regulation that the of the element that within the region of the element in regulation of the amastin this we the of the region the region this of the of in the the of regulation the element in of nucleotides of of the region in regulation the regulatory of the amastin region of in the region the element in the Leishmania conserved in Leishmania amastin gene is that a within the region regulation that this in several mRNAs. within this region this in of the of luciferase reporter we that within the amastin 3′-UTR region the of in the of the regulation in of these regulatory the of the 3′-UTR stage-specific we of the in in the the the in that of the 3′-UTR that the regulation these that the of 3′-UTR identified within the region of the amastin is in is the of that amastin in Leishmania in is that is the nucleotides of the amastin we the that the within the 770 nucleotides of the 3′-UTR a in this the of the the of the amastin that in we a of of the regulatory 3′-UTR gene the of Leishmania the gene the region the the region the element the the a Leishmania is of of in the the of the amastin in these we that the in amastin in the the the luciferase reporter the of the 3′-UTR the of a of these in the 3′-UTR highly we the of 3′-UTR regulatory of the amastin the element element the that a at the these a the of the amastin 3′-UTR regulatory that the the a a of of the in the the the is the at in a in the of the the of the that is the of the 3′-UTR regulatory is in in the in of the amastin 3′-UTR regulatory in of a the a at the the of the the of the in the of a the of the of in the of in the of the of the of in a a of the in a of the regulation of stage-specific gene expression in Leishmania is in the in that 3′-UTR a in the of in Leishmania is a of that the of the of of the of the amastin in we the of Leishmania the gene the amastin the 3′-UTR regulatory Leishmania the the in the of these the of amastin that of the amastin in the regulation of the amastin in the of the in the amastin the of the amastin 3′-UTR the the of the regulation of the amastin using a luciferase reporter the amastin the in the luciferase the of of a the the of Leishmania in the in the of is the of the we these regulation of amastin mRNAs. of the 3′-UTR regulatory in of the the of of the in within in the the at the of the these we that regulation is Leishmania the the of Leishmania within that amastin translation. regulation of the amastin the the the a of the of these the luciferase in a at at the of amastin a that is of the the the in the of the amastin the of of the that the of amastin in the Here we that 3′-UTR that amastin in in these of of the amastin that is a 2055-nt 3′-UTR in that stage-specific amastin gene expression of that within 3′-UTR of the recently characterized a large family of developmentally regulated in surface of the amastin gene that we in we identified a region of 770 in the 3′-UTR that regulates stage-specific gene expression at the level of the nucleotides of this regulatory region conserved among several in the of the amastin the of the in several of that of the element We that this conserved element the regulation of the amastin this we a of within the region that we of the luciferase reporter gene Leishmania of expression in in using in the region in in element in of the Leishmania the of the amastin gene that the region of a nucleotides of of the element this we a that of of the of the regulation the region the nucleotides that the element is regulation that the of the element that within the region of the element in regulation of the amastin this we the of the region the region this of the of in the the of regulation the element in of nucleotides of of the region in regulation the regulatory of the amastin region of in the region the element in the Leishmania conserved in Leishmania amastin gene is that a within the region regulation that this in several mRNAs. within this region this in of the of luciferase reporter we that within the amastin 3′-UTR region the of in the of the regulation in of these regulatory the of the 3′-UTR stage-specific we of the in in the the the in that of the 3′-UTR that the regulation these that the of 3′-UTR identified within the region of the amastin is in is the of that amastin in Leishmania in is that is the nucleotides of the amastin we the that the within the 770 nucleotides of the 3′-UTR a in this the of the the of the amastin that in we a of of the regulatory 3′-UTR gene the of Leishmania the gene the region the the region the element the the a Leishmania is of of in the the of the amastin in these we that the in amastin in the the the luciferase reporter the of the 3′-UTR the of a of these in the 3′-UTR highly we the of 3′-UTR regulatory of the amastin the element element the that a at the these a the of the amastin 3′-UTR regulatory that the the a a of of the in the the the is the at in a in the of the the of the that is the of the 3′-UTR regulatory is in in the in of the regulation of stage-specific gene expression in Leishmania is in the in that 3′-UTR a in the of in Leishmania is a of that the of the of of the of the amastin in we the of Leishmania the gene the amastin the 3′-UTR regulatory Leishmania the the in the of these the of amastin that of the amastin in the of Leishmania in the in the of is the of the we these regulation of amastin mRNAs. of the 3′-UTR regulatory in of the the of of the in within in the the at the of the these we that regulation is Leishmania the that the of amastin in the Here we that 3′-UTR that amastin in in these of of the amastin that is a 2055-nt 3′-UTR in that stage-specific amastin gene expression of that within we a of regulation that 3′-UTR of the developmentally regulated amastin in We these of the within is gene regulation in this the that stage-specific regulation of gene expression in that regulation of the amastin is the 770 nucleotides of the a ∼450-nt element that is highly conserved among several Leishmania a of We that this 3′-UTR element in stage-specific Here we that the element of the amastin in the the that of the of the the this element is a of the the of is that the element that the of the the in this that regulation the conserved element the region of a of of of the that is that of the the of in of these the the presence of that conserved among the of the element the that several Leishmania regulated a of in of regulation a the a within is that the regulation is a the is the the of that stage-specific regulation in is that amastin is 3′-UTR in the a region of that is in of the amastin is in the this identified region in of the regulatory element the a of this in is that the of in that in of the amastin at regulation at the level of a the of the amastin surface the of the in the presence of a that of translation. the of the 3′-UTR in the the that regulation these at the level of is the of regulation in is the of in of 3′-UTR of several of in of the of the of a a at the of the 3′-UTR in identified the the in this is that regulation of the amastin gene family is at the level of at the level of that within is that the of regulation that the 3′-UTR regulatory of amastin mRNAs. system of we that of these 3′-UTR regulatory a in amastin regulation that the of amastin in the of the that this is in the of the in amastin is highly the the in the that translation. in that in of of that a of in in the in this a of regulation the developmentally regulated amastin gene family in Leishmania that is in within a regulation that is in a regulatory element within the is in is the that the of regulation that stage-specific gene expression in the of these gene regulation in of in the in a in within the 3′-UTR the in in in a in within the 3′-UTR the in in in the of in in within the 3′-UTR the in in in a in within the 3′-UTR the in in in in in a Here we a of regulation that 3′-UTR of the developmentally regulated amastin in We these of the within is gene regulation in this the that stage-specific regulation of gene expression in We that regulation of the amastin is the 770 nucleotides of the a ∼450-nt element that is highly conserved among several Leishmania a of We that this 3′-UTR element in stage-specific Here we that the element of the amastin in the the that of the of the the this element is a of the the of is that the element that the of the the in this that regulation the conserved element the region of a of of of the that is that of the the of in of these the the presence of that conserved among the of the element the that several Leishmania regulated a of in of regulation a the a within is that the regulation is a the is the the of that stage-specific regulation in is that amastin is 3′-UTR in the a region of that is in of the amastin is in the this identified region in of the regulatory element the a of this in is that the of in that in of the amastin at regulation at the level of a the of the amastin surface the of the in the presence of a that of translation. the of the 3′-UTR in the the that regulation these at the level of is the of regulation in is the of in of 3′-UTR of several of in of the of the of a a at the of the 3′-UTR in identified the the in this is that regulation of the amastin gene family is at the level of at the level of that within is that the of regulation that the 3′-UTR regulatory of amastin mRNAs. system of we that of these 3′-UTR regulatory a in amastin regulation that the of amastin in the of the that this is in the of the in amastin is highly the the in the that translation. in that in of of that a of in in the in this a of regulation the developmentally regulated amastin gene family in Leishmania that is in within a regulation that is in a regulatory element within the is in is the that the of regulation that stage-specific gene expression in the of these We of the We

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