Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic>
ABSTRACT During their parasitic life cycle, through sandflies and vertebrate hosts, Leishmania parasites confront strikingly different environments, including abrupt changes in pH and temperature, to which they must rapidly adapt. These adaptations include alterations in Leishmania gene expression,...
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American Society for Microbiology
2019
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oai:doaj.org-article:0401eec1aa5c46ef927e63e93675b9532021-11-15T15:22:22ZDisruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic>10.1128/mSphere.00644-182379-5042https://doaj.org/article/0401eec1aa5c46ef927e63e93675b9532019-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00644-18https://doaj.org/toc/2379-5042ABSTRACT During their parasitic life cycle, through sandflies and vertebrate hosts, Leishmania parasites confront strikingly different environments, including abrupt changes in pH and temperature, to which they must rapidly adapt. These adaptations include alterations in Leishmania gene expression, metabolism, and morphology, allowing them to thrive as promastigotes in the sandfly and as intracellular amastigotes in the vertebrate host. A critical aspect of Leishmania metabolic adaptation to these changes is maintenance of efficient mitochondrial function in the hostile vertebrate environment. Such functions, including generation of ATP, depend upon the expression of many mitochondrial proteins, including subunits of cytochrome c oxidase (COX). Significantly, under mammalian temperature conditions, expression of Leishmania major COX subunit IV (LmCOX4) and virulence are dependent upon two copies of LACK, a gene that encodes the ribosome-associated scaffold protein, LACK (Leishmania ortholog of RACK1 [receptor for activated C kinase]). Targeted replacement of an endogenous LACK copy with a putative ribosome-binding motif-disrupted variant (LACKR34D35G36→LACKD34D35E36) resulted in thermosensitive parasites that showed diminished LmCOX4 expression, mitochondrial fitness, and replication in macrophages. Surprisingly, despite these phenotypes, LACKD34D35E36 associated with monosomes and polysomes and showed no major impairment of global protein synthesis. Collectively, these data suggest that wild-type (WT) LACK orchestrates robust LmCOX4 expression and mitochondrial fitness to ensure parasite virulence, via optimized functional interactions with the ribosome. IMPORTANCE Leishmania parasites are trypanosomatid protozoans that persist in infected human hosts to cause a spectrum of pathologies, from cutaneous and mucocutaneous manifestations to visceral leishmaniasis caused by Leishmania donovani. The latter is usually fatal if not treated. Persistence of L. major in the mammalian host depends upon maintaining gene-regulatory programs to support essential parasite metabolic functions. These include expression and assembly of mitochondrial L. major cytochrome c oxidase (LmCOX) subunits, important for Leishmania ATP production. Significantly, under mammalian conditions, WT levels of LmCOX subunits require threshold levels of the Leishmania ribosome-associated scaffold protein, LACK. Unexpectedly, we find that although disruption of LACK’s putative ribosome-binding motif does not grossly perturb ribosome association or global protein synthesis, it nonetheless impairs COX subunit expression, mitochondrial function, and virulence. Our data indicate that the quality of LACK’s interaction with Leishmania ribosomes is critical for LmCOX subunit expression and parasite mitochondrial function in the mammalian host. Collectively, these findings validate LACK’s ribosomal interactions as a potential therapeutic target.Daviel CardenasCharity SylvesterBo CaoCatherine S. NationJuan C. PizarroHua LuJessie GuidryEdward J. WojcikBen L. KellyAmerican Society for MicrobiologyarticleLACKLeishmaniaRACK1cytochrome c oxidasemitochondriaparasiteMicrobiologyQR1-502ENmSphere, Vol 4, Iss 2 (2019) |
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DOAJ |
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EN |
| topic |
LACK Leishmania RACK1 cytochrome c oxidase mitochondria parasite Microbiology QR1-502 |
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LACK Leishmania RACK1 cytochrome c oxidase mitochondria parasite Microbiology QR1-502 Daviel Cardenas Charity Sylvester Bo Cao Catherine S. Nation Juan C. Pizarro Hua Lu Jessie Guidry Edward J. Wojcik Ben L. Kelly Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| description |
ABSTRACT During their parasitic life cycle, through sandflies and vertebrate hosts, Leishmania parasites confront strikingly different environments, including abrupt changes in pH and temperature, to which they must rapidly adapt. These adaptations include alterations in Leishmania gene expression, metabolism, and morphology, allowing them to thrive as promastigotes in the sandfly and as intracellular amastigotes in the vertebrate host. A critical aspect of Leishmania metabolic adaptation to these changes is maintenance of efficient mitochondrial function in the hostile vertebrate environment. Such functions, including generation of ATP, depend upon the expression of many mitochondrial proteins, including subunits of cytochrome c oxidase (COX). Significantly, under mammalian temperature conditions, expression of Leishmania major COX subunit IV (LmCOX4) and virulence are dependent upon two copies of LACK, a gene that encodes the ribosome-associated scaffold protein, LACK (Leishmania ortholog of RACK1 [receptor for activated C kinase]). Targeted replacement of an endogenous LACK copy with a putative ribosome-binding motif-disrupted variant (LACKR34D35G36→LACKD34D35E36) resulted in thermosensitive parasites that showed diminished LmCOX4 expression, mitochondrial fitness, and replication in macrophages. Surprisingly, despite these phenotypes, LACKD34D35E36 associated with monosomes and polysomes and showed no major impairment of global protein synthesis. Collectively, these data suggest that wild-type (WT) LACK orchestrates robust LmCOX4 expression and mitochondrial fitness to ensure parasite virulence, via optimized functional interactions with the ribosome. IMPORTANCE Leishmania parasites are trypanosomatid protozoans that persist in infected human hosts to cause a spectrum of pathologies, from cutaneous and mucocutaneous manifestations to visceral leishmaniasis caused by Leishmania donovani. The latter is usually fatal if not treated. Persistence of L. major in the mammalian host depends upon maintaining gene-regulatory programs to support essential parasite metabolic functions. These include expression and assembly of mitochondrial L. major cytochrome c oxidase (LmCOX) subunits, important for Leishmania ATP production. Significantly, under mammalian conditions, WT levels of LmCOX subunits require threshold levels of the Leishmania ribosome-associated scaffold protein, LACK. Unexpectedly, we find that although disruption of LACK’s putative ribosome-binding motif does not grossly perturb ribosome association or global protein synthesis, it nonetheless impairs COX subunit expression, mitochondrial function, and virulence. Our data indicate that the quality of LACK’s interaction with Leishmania ribosomes is critical for LmCOX subunit expression and parasite mitochondrial function in the mammalian host. Collectively, these findings validate LACK’s ribosomal interactions as a potential therapeutic target. |
| format |
article |
| author |
Daviel Cardenas Charity Sylvester Bo Cao Catherine S. Nation Juan C. Pizarro Hua Lu Jessie Guidry Edward J. Wojcik Ben L. Kelly |
| author_facet |
Daviel Cardenas Charity Sylvester Bo Cao Catherine S. Nation Juan C. Pizarro Hua Lu Jessie Guidry Edward J. Wojcik Ben L. Kelly |
| author_sort |
Daviel Cardenas |
| title |
Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| title_short |
Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| title_full |
Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| title_fullStr |
Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| title_full_unstemmed |
Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome <italic toggle="yes">c</italic> Oxidase Subunit Expression in <italic toggle="yes">Leishmania major</italic> |
| title_sort |
disruption of the putative ribosome-binding motif of a scaffold protein impairs cytochrome <italic toggle="yes">c</italic> oxidase subunit expression in <italic toggle="yes">leishmania major</italic> |
| publisher |
American Society for Microbiology |
| publishDate |
2019 |
| url |
https://doaj.org/article/0401eec1aa5c46ef927e63e93675b953 |
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