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

• Published in: Nature communications Vol. 12; no. 1; pp. 1244 - 15
• Main Authors: 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.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 23.02.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/109; 45/41; 631/326/417/1716; 631/326/417/2549; Animals; Cell Differentiation; Cell Survival; Clustering; Colonization; CRISPR-Associated Protein 9 - metabolism; CRISPR-Cas Systems - genetics; Deletion mutant; Differentiation; Disease transmission; Female; Flagella - enzymology; Functional analysis; Gene Deletion; Genes; Humanities and Social Sciences; Interrogation; Intracellular; Intracellular signalling; Kinases; Leishmania; Leishmania mexicana; Leishmania mexicana - cytology; Leishmania mexicana - enzymology; Leishmaniasis - parasitology; Leishmaniasis - pathology; Life cycles; Macrophages; Mice; Mice, Inbred BALB C; Models, Biological; multidisciplinary; Mutation - genetics; Parasites; Parasitism; Phenotyping; Promastigotes; Protein kinase; Protein Kinases - genetics; Protein Kinases - metabolism; Proteins; Proteome - metabolism; Protozoa; Psychodidae - parasitology; Regulators; Science; Science (multidisciplinary); Signal transduction; Signaling; Survival
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-21360-8

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. Protein kinases are fundamental in cellular signalling required for Leishmania survival throughout the life cycle. Here, Baker and Catta-Preta et al. report on a kinome-wide functional study in Leishmania mexicana to define protein kinases with roles in life cycle transition.