Mitochondrial associated ubiquitin fold modifier-1 mediated protein conjugation in Leishmania donovani.
In this report, we demonstrate the existence of the ubiquitin fold modifier-1 (Ufm1) and its conjugation pathway in trypanosomatid parasite Leishmania donovani. LdUfm1 is activated by E1-like enzyme LdUba5. LdUfc1 (E2) specifically interacted with LdUfm1 and LdUba5 to conjugate LdUfm1 to proteinaceous targets. Mass spectrometry analysis revealed that LdUfm1 is conjugated to Leishmania protein targets that are associated with mitochondria. Immunofluorescence experiments showed that Leishmania Ufm1, Uba5 and Ufc1 are associated with the mitochondria. The demonstration that all the components of this system as well as the substrates are associated with mitochondrion suggests it may have physiological roles not yet described in any other organism. Overexpression of a non-conjugatable form of LdUfm1 and an active site mutant of LdUba5 resulted in reduced survival of Leishmania in the macrophage. Since mitochondrial activities are developmentally regulated in the life cycle of trypanosomatids, Ufm1 mediated modifications of mitochondrial proteins may be important in such regulation. Thus, Ufm1 conjugation pathway in Leishmania could be explored as a potential drug target in the control of Leishmaniasis.
Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Bethesda, Maryland, United States of America.
Sreenivas
S
Gannavaram
Paresh
P
Sharma
Robert C
RC
Duncan
Poonam
P
Salotra
Hira L
HL
Nakhasi
D006801:Humans; D007893:Leishmania donovani / Q000737:chemistry* / Q000378:metabolism; D008264:Macrophages / Q000469:parasitology; D013058:Mass Spectrometry; D008928:Mitochondria / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D044764:Ubiquitin-Activating Enzymes
D015800:Protozoan Proteins; D044764:Ubiquitin-Activating Enzymes
EMBO J. 2006 Aug 9;25(15):3618-26
16874301
Biochem Soc Trans. 2010 Feb;38(Pt 1):132-6
20074047
EMBO J. 2004 May 5;23(9):1977-86
15071506
Am J Trop Med Hyg. 1993 Nov;49(5):545-51
8250094
Annu Rev Cell Dev Biol. 2006;22:159-80
16753028
Ann N Y Acad Sci. 2008 Dec;1147:242-53
19076446
Curr Opin Microbiol. 2007 Dec;10(6):569-77
18177626
Gene. 1994 Jun 24;144(1):45-51
8026757
Mol Cell. 2007 Jun 22;26(6):891-8
17588522
J Cell Sci. 2008 Jan 1;121(Pt 1):99-109
18073240
Biol Res. 1993;26(1-2):151-7
7670528
J Biol Chem. 1991 Nov 5;266(31):21018-24
1657944
Parasitology. 2007 Oct;134(Pt 11):1527-39
17553180
J Biol Chem. 2007 Feb 23;282(8):5256-62
17182609
Mol Biochem Parasitol. 2006 Jul;148(1):69-78
16600399
Parasitology. 2006;133 Suppl:S87-112
17274851
Annu Rev Pharmacol Toxicol. 2006;46:189-213
16402903
Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5542-6
1376922
Mol Cell. 2009 Feb 13;33(3):275-86
19217402
EMBO Rep. 2008 Mar;9(3):280-6
18274552
EMBO Rep. 2009 Jul;10(7):748-54
19407830
PLoS One. 2007 Oct 24;2(10):e1066
17957250
Mol Microbiol. 2006 Sep;61(5):1187-95
16925553
Mol Biochem Parasitol. 1996 Apr;77(1):57-64
8784772
Biochim Biophys Acta. 2008 Nov;1784(11):1742-9
18640292
Cell Microbiol. 2008 Oct;10(10):1999-2011
18547337
Mol Biochem Parasitol. 1996 Jun;78(1-2):79-90
8813679
J Biol Chem. 1988 Sep 5;263(25):12698-704
2457589
Mol Biochem Parasitol. 2000 Jan 5;105(1):115-25
10613704
Exp Cell Res. 2010 Mar 10;316(5):704-15
20045687
Cell Microbiol. 2010 Jan;12(1):19-30
19702650
Future Oncol. 2007 Apr;3(2):191-9
17381419
Eukaryot Cell. 2007 Nov;6(11):1943-52
17905920
BMC Biochem. 2007 Nov 22;8 Suppl 1:S14
18047738
Mol Cell Proteomics. 2008 Mar;7(3):534-45
18073385
Cell Death Differ. 2010 Jan;17(1):4-13
19557013
Science. 2007 Jan 12;315(5809):201-5
17218518
Arch Med Res. 2006 Jul;37(5):593-601
16740428
Biochemistry. 2001 Jan 30;40(4):1053-62
11170428
Traffic. 2008 Sep;9(10):1681-97
18657071
J Biol Chem. 2010 Feb 19;285(8):5417-27
20018847
IUBMB Life. 2010 Feb;62(2):140-57
20073038
Annu Rev Biochem. 1998;67:425-79
9759494
Nature. 2009 Mar 26;458(7237):422-9
19325621