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Mol Cell Proteomics. 2019 Jun;18(6):1171-1182. doi: 10.1074/mcp.RA119.001299. Epub 2019 Mar 28.

Proteomics of Asrij Perturbation in Drosophila Lymph Glands for Identification of New Regulators of Hematopoiesis.

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From the ‡Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India.
§Institute of Bioinformatics, International Technology Park, Bangalore 560066, India.
¶Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA.
‖NIMHANS-IOB Proteomics and Bioinformatics Laboratory, Neurobiology Research Centre, National Institute of Mental Health and Neurosciences, Bangalore 560029, India.
**Center for Systems Biology and Molecular Medicine, Yenepoya Research Center, Yenepoya (Deemed to be University), Mangalore-575018, India.
From the ‡Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India;
‡‡Institute for Stem Cell Biology and Regenerative Medicine, GKVK, Bellary Road, Bangalore 560065, India.


Hematopoiesis is the process of differentiation of precursor blood cells into mature blood cells that is controlled by a complex set of molecular interactions. Understanding hematopoiesis is important for the study of hematological disorders. However, a comprehensive understanding of how physiological and genetic mechanisms regulate blood cell precursor maintenance and differentiation is lacking. Owing to simplicity and ease of genetic analysis, the Drosophila melanogaster lymph gland (LG) is an excellent model to study hematopoiesis. Here, we quantitatively analyzed the LG proteome under genetic conditions that either maintain precursors or promote their differentiation in vivo, by perturbing expression of Asrij, a conserved endosomal regulator of hematopoiesis. Using iTRAQ-based quantitative proteomics, we determined the relative expression levels of proteins in Asrij-knockout and overexpressing LGs from 1500 larval dissections compared with wild type. Our data showed that at least 6.5% of the Drosophila proteome is expressed in wild type LGs. Of the 2133 proteins identified, 780 and 208 proteins were common to previously reported cardiac tube and hemolymph proteomes, respectively, resulting in the identification of 1238 proteins exclusive to the LG. Perturbation of Asrij levels led to differential expression of 619 proteins, of which 27% have human homologs implicated in various diseases. Proteins regulating metabolism, immune system, signal transduction and vesicle-mediated transport were significantly enriched. Immunostaining of representative candidates from the enriched categories and previous reports confirmed 73% of our results, indicating the validity of our LG proteome. Our study provides, for the first time, an in vivo proteomics resource for identifying novel regulators of hematopoiesis that will also be applicable to understanding vertebrate blood cell development.


Asrij; Blood*; Differentiation*; Drosophila melanogaster*; Knockouts*; Mass Spectrometry; hematopoiesis; iTRAQ; lymph gland; proteome

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