| Full text | |
| Author(s): Show less - |
Morea, Edna G. O.
[1]
;
Viviescas, Maria Alejandra
[1]
;
Fernandes, Carlos A. H.
[2]
;
Matioli, Fabio F.
[2]
;
Lira, Cristina B. B.
[1]
;
Fernandez, Maribel F.
[3]
;
Moraes, Barbara S.
[4]
;
da Silva, Marcelo S.
[5]
;
Storti, Camila B.
[1]
;
Fontes, Marcos R. M.
[2]
;
Cano, Maria Isabel N.
[1]
Total Authors: 11
|
| Affiliation: | [1] Sao Paulo State Univ UNESP, Biosci Inst, Genet Dept, BR-18618689 Botucatu, SP - Brazil
[2] Sao Paulo State Univ (UNESP), Biosci Inst, Biophys & Phys Dept, Botucatu, SP - Brazil
[3] ITPAC Porto Nacl SA, Inst Tocantinense Presidente Antonio Carlos LTDA, Porto Nacional, TO - Brazil
[4] PROAHSA Programa Estudos Avancados Adm Hosp & Sis, Sao Paulo - Brazil
[5] Butantan Inst, Ctr Toxins Immune Response & Cell Signaling CeTIC, Lab Especial Ciclo Celular, Sao Paulo, SP - Brazil
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1861, n. 11, A, p. 2583-2597, NOV 2017. |
| Web of Science Citations: | 1 |
| Abstract | |
Background: Leishmania spp. telomeres are composed of 5'-TTAGGG-3' repeats associated with proteins. We have previously identified LaRbp38 and LaRPA-1 as proteins that bind the G-rich telomeric strand. At that time, we had also partially characterized a protein: DNA complex, named LaGT1, but we could not identify its protein component. Methods and results: Using protein-DNA interaction and competition assays, we confirmed that LaGT1 is highly specific to the G-rich telomeric single-stranded DNA. Three protein bands, with LaGT1 activity, were isolated from affinity-purified protein extracts in-gel digested, and sequenced de novo using mass spectrometry analysis. In silico analysis of the digested peptide identified them as a putative calmodulin with sequences identical to the T. cruzi calmodulin. In the Leishmania genome, the calmodulin ortholog is present in three identical copies. We cloned and sequenced one of the gene copies, named it LCalA, and obtained the recombinant protein. Multiple sequence alignment and molecular modeling showed that LCalA shares homology to most eukaryotes calmodulin. In addition, we demonstrated that LCalA is nuclear, partially co-localizes with telomeres and binds in vivo the G-rich telomeric strand. Recombinant LCalA can bind specifically and with relative affinity to the G-rich telomeric single-strand and to a 3'G-overhang, and DNA binding is calcium dependent. Conclusions: We have described a novel candidate component of Leishmania telomeres, LCalA, a nuclear calmodulin that binds the G-rich telomeric strand with high specificity and relative affinity, in a calcium-dependent manner. (AU) | |
| FAPESP's process: | 15/18641-8 - Caracterization of long-noncoding telomeric RNAs in Leishmania major |
| Grantee: | Maria Isabel Nogueira Cano |
| Support type: | Regular Research Grants |
| FAPESP's process: | 12/50161-8 - Study of the structure and function of the Hsp90 chaperone with emphasis on its role in cellular homeostasis |
| Grantee: | Carlos Henrique Inacio Ramos |
| Support type: | Research Projects - Thematic Grants |