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The natural absence of RPA1N domain did not impair Leishmania amazonensis RPA-1 participation in DNA damage response and telomere protection

Published online by Cambridge University Press:  07 February 2013

RITA DE CÁSSIA VIVEIROS DA SILVEIRA ,
MARCELO SANTOS DA SILVA ,
VINÍCIUS SANTANA NUNES ,
ARINA MARINA PEREZ  and
MARIA ISABEL NOGUEIRA CANO
RITA DE CÁSSIA VIVEIROS DA SILVEIRA
Affiliation:
Departamento de Genética, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Botucatu, SP, 18618-970, Brazil
MARCELO SANTOS DA SILVA
Affiliation:
Departamento de Genética, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Botucatu, SP, 18618-970, Brazil
VINÍCIUS SANTANA NUNES
Affiliation:
Departamento de Genética, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Botucatu, SP, 18618-970, Brazil
ARINA MARINA PEREZ
Affiliation:
Departamento de Genética, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Botucatu, SP, 18618-970, Brazil
MARIA ISABEL NOGUEIRA CANO*
Affiliation:
Departamento de Genética, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Botucatu, SP, 18618-970, Brazil
*
*Corresponding author. E-mail: [email protected]
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Summary

We have previously shown that the subunit 1 of Leishmania amazonensis RPA (LaRPA-1) alone binds the G-rich telomeric strand and is structurally different from other RPA-1. It is analogous to telomere end-binding proteins described in model eukaryotes whose homologues were not identified in the protozoan´s genome. Here we show that LaRPA-1 is involved with damage response and telomere protection although it lacks the RPA1N domain involved with the binding with multiple checkpoint proteins. We induced DNA double-strand breaks (DSBs) in Leishmania using phleomycin. Damage was confirmed by TUNEL-positive nuclei and triggered a G1/S cell cycle arrest that was accompanied by nuclear accumulation of LaRPA-1 and RAD51 in the S phase of hydroxyurea-synchronized parasites. DSBs also increased the levels of RAD51 in non-synchronized parasites and of LaRPA-1 and RAD51 in the S phase of synchronized cells. More LaRPA-1 appeared immunoprecipitating telomeres in vivo and associated in a complex containing RAD51, although this interaction needs more investigation. RAD51 apparently co-localized with few telomeric clusters but it did not immunoprecipitate telomeric DNA. These findings suggest that LaRPA-1 and RAD51 work together in response to DNA DSBs and at telomeres, upon damage, LaRPA-1 works probably to prevent loss of single-stranded DNA and to assume a capping function.

Keywords

Leishmania amazonensisDNA double-stranded breakLaRPA-1RAD51telomeres
Type
Research Article
Information
Parasitology , Volume 140 , Issue 4 , April 2013 , pp. 547 - 559
Copyright
Copyright © Cambridge University Press 2013

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