Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

N Baker; C M C Catta-Preta; R Neish; J Sadlova; B Powell; E V C Alves-Ferreira; V Geoghegan; J B T Carnielli; K Newling; C Hughes; B Vojtkova; J Anand; A Mihut; P B Walrad; L G Wilson; J W Pitchford; P Volf; J C Mottram

doi:10.1038/s41467-021-21360-8

Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

N Baker, C M C Catta-Preta, R Neish, J Sadlova, B Powell, E V C Alves-Ferreira, V Geoghegan, J B T Carnielli, K Newling, C Hughes, B Vojtkova, J Anand, A Mihut, P B Walrad, L G Wilson, J W Pitchford, P Volf, J C Mottram

Research output: Contribution to journal › Article › peer-review

Abstract

Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.

Original language	English
Article number	1244
Number of pages	15
Journal	Nature Communications
Volume	12
Issue number	1
Early online date	23 Feb 2021
DOIs	https://doi.org/10.1038/s41467-021-21360-8 https://doi.org/10.1038/s41467-021-21360-8
Publication status	Published - 23 Feb 2021

Bibliographical note

Keywords

Animals
CRISPR-Associated Protein 9/metabolism
CRISPR-Cas Systems/genetics
Cell Differentiation
Cell Survival
Female
Flagella/enzymology
Gene Deletion
Leishmania mexicana/cytology
Leishmaniasis/parasitology
Mice, Inbred BALB C
Models, Biological
Mutation/genetics
Protein Kinases/genetics
Proteome/metabolism
Psychodidae/parasitology

Access to Document

s41467-021-21360-8
© The Author(s) 2021
Final published version, 2.93 MBLicence: CC BY

Cite this

Baker, N., Catta-Preta, C. M. C., Neish, R., Sadlova, J., Powell, B., Alves-Ferreira, E. V. C., Geoghegan, V., Carnielli, J. B. T., Newling, K., Hughes, C., Vojtkova, B., Anand, J., Mihut, A., Walrad, P. B., Wilson, L. G., Pitchford, J. W., Volf, P., & Mottram, J. C. (2021). Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival. Nature Communications, 12(1), Article 1244. https://doi.org/10.1038/s41467-021-21360-8, https://doi.org/10.1038/s41467-021-21360-8

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abstract = "Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.",

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Baker, N, Catta-Preta, CMC , Neish, R, Sadlova, J, Powell, B, Alves-Ferreira, EVC, Geoghegan, V, Carnielli, JBT, Newling, K , Hughes, C, Vojtkova, B, Anand, J, Mihut, A, Walrad, PB , Wilson, LG , Pitchford, JW, Volf, P & Mottram, JC 2021, 'Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival', Nature Communications, vol. 12, no. 1, 1244. https://doi.org/10.1038/s41467-021-21360-8, https://doi.org/10.1038/s41467-021-21360-8

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T1 - Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

AU - Baker, N

AU - Catta-Preta, C M C

AU - Neish, R

AU - Sadlova, J

AU - Powell, B

AU - Alves-Ferreira, E V C

AU - Geoghegan, V

AU - Carnielli, J B T

AU - Newling, K

AU - Hughes, C

AU - Vojtkova, B

AU - Anand, J

AU - Mihut, A

AU - Walrad, P B

AU - Wilson, L G

AU - Pitchford, J W

AU - Volf, P

AU - Mottram, J C

N1 - © The Author(s) 2021

PY - 2021/2/23

Y1 - 2021/2/23

N2 - Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.

AB - Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.

KW - Animals

KW - CRISPR-Associated Protein 9/metabolism

KW - CRISPR-Cas Systems/genetics

KW - Cell Differentiation

KW - Cell Survival

KW - Female

KW - Flagella/enzymology

KW - Gene Deletion

KW - Leishmania mexicana/cytology

KW - Leishmaniasis/parasitology

KW - Mice, Inbred BALB C

KW - Models, Biological

KW - Mutation/genetics

KW - Protein Kinases/genetics

KW - Proteome/metabolism

KW - Psychodidae/parasitology

U2 - 10.1038/s41467-021-21360-8

DO - 10.1038/s41467-021-21360-8

M3 - Article

C2 - 33623024

SN - 2041-1723

VL - 12

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 1244

ER -