LaRbp38: a Leishmania amazonensis protein that binds nuclear and kinetoplast DNAs.
Leishmania amazonensis causes a wide spectrum of leishmaniasis. There are no vaccines or adequate treatment for leishmaniasis, therefore there is considerable interest in the identification of new targets for anti-leishmania drugs. The central role of telomere-binding proteins in cell maintenance makes these proteins potential targets for new drugs. In this work, we used a combination of purification chromatographies to screen L. amazonensis proteins for molecules capable of binding double-stranded telomeric DNA. This approach resulted in the purification of a 38kDa polypeptide that was identified by mass spectrometry as Rbp38, a trypanosomatid protein previously shown to stabilize mitochondrial RNA and to associate with nuclear and kinetoplast DNAs. Western blotting and supershift assays confirmed the identity of the protein as LaRbp38. Competition and chromatin immunoprecipitation assays confirmed that LaRbp38 interacted with kinetoplast and nuclear DNAs in vivo and suggested that LaRbp38 may have dual cellular localization and more than one function.
Lira
C B B
CB
Departamento de Genética, IB, Universidade Estadual de São Paulo, UNESP, 18618-000, Botucatu, SP, Brazil.
Siqueira Neto
J L
JL
Giardini
M A
MA
Winck
F V
FV
Ramos
C H I
CH
Cano
M I N
MI
D000818:Animals; D000977:Antiparasitic Agents / Q000494:pharmacology; D001667:Binding, Competitive; D002467:Cell Nucleus / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D004247:DNA / Q000378:metabolism; D018105:DNA, Kinetoplast / Q000737:chemistry / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D047468:Immunoprecipitation; D007891:Leishmania / Q000378:metabolism; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D012313:RNA / Q000737:chemistry; D000077278:RNA, Mitochondrial; D016615:Telomere / Q000737:chemistry / Q000648:ultrastructure
D000977:Antiparasitic Agents; D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins; D010455:Peptides; D000077278:RNA, Mitochondrial; D012313:RNA; D004247:DNA