caa a22 4500 606233784 CHVBK 20210128101234.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s11010-015-2366-6 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2366-6 A unique, highly conserved secretory invertase is differentially expressed by promastigote developmental forms of all species of the human pathogen, Leishmania [Elektronische Daten] [Todd Lyda, Manju Joshi, John Andersen, Andrew Kelada, Joshua Owings, Paul Bates, Dennis Dwyer] Leishmania are protozoan pathogens of humans that exist as extracellular promastigotes in the gut of their sand fly vectors and as obligate intracellular amastigotes within phagolysosomes of infected macrophages. Between infectious blood meal feeds, sand flies take plant juice meals that contain sucrose and store these sugars in their crop. Such sugars are regurgitated into the sand fly anterior midgut where they impact the developing promastigote parasite population. In this report we showed that promastigotes of all Leishmania species secreted an invertase/sucrase enzyme during their growth in vitro. In contrast, neither L. donovani nor L. mexicana amastigotes possessed any detectable invertase activity. Importantly, no released/secreted invertase activity was detected in culture supernatants from either Trypanosoma brucei or Trypanosoma cruzi. Using HPLC, the L. donovani secretory invertase was isolated and subjected to amino acid sequencing. Subsequently, we used a molecular approach to identify the LdINV and LmexINV genes encoding the ~72kDa invertases produced by these organisms. Interestingly, we identified high fidelity LdINV-like homologs in the genomes of all Leishmania sp. but none were present in either T. brucei or T. cruzi. Northern blot and RT-PCR analyses showed that these genes were developmentally/differentially expressed in promastigotes but not amastigotes of these parasites. Homologous transfection studies demonstrated that these genes in fact encoded the functional secretory invertases produced by these parasites. Cumulatively, our results suggest that these secretory enzymes play critical roles in the survival/growth/development and transmission of all Leishmania parasites within their sand fly vector hosts. Springer Science+Business Media New York (outside the USA), 2015 Leishmania nationallicence Protozoan pathogen nationallicence Human parasite nationallicence Invertase nationallicence Secretory enzyme nationallicence Lyda Todd 105 Collings Street BRC 216, 29634, Clemson, SC, USA aut Joshi Manju Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut Andersen John Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, USA aut Kelada Andrew Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut Owings Joshua Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut Bates Paul Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, LA1 4YQ, Lancaster, UK aut Dwyer Dennis Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 404/1-2(2015-06-01), 53-77 0300-8177 404:1-2<53 2015 404 11010 https://doi.org/10.1007/s11010-015-2366-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2366-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lyda Todd 105 Collings Street BRC 216, 29634, Clemson, SC, USA aut NATIONALLICENCE 700 1- Joshi Manju Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Andersen John Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, USA aut NATIONALLICENCE 700 1- Kelada Andrew Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Owings Joshua Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Bates Paul Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, LA1 4YQ, Lancaster, UK aut NATIONALLICENCE 700 1- Dwyer Dennis Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892-0425, Bethesda, MD, USA aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 404/1-2(2015-06-01), 53-77 0300-8177 404:1-2<53 2015 404 11010