| Full text | |
| Author(s): |
Balasco Serrao, Vitor Hugo
[1, 2]
;
de Freitas Fernandes, Adriano
[1]
;
Mansor Basso, Luis Guilherme
[3, 4]
;
Scortecci, Jessica Fernandes
[1, 5]
;
Crusca Junior, Edson
[6]
;
Lopes Cornelio, Marinonio
[7]
;
Monson de Souza, Bibiana
[8]
;
Palma, Mario Sergio
[8]
;
de Oliveira Neto, Mario
[9]
;
Thiemann, Otavio Henrique
[1, 10]
Total Authors: 10
|
| Affiliation: | [1] Univ Sao Paulo, Phys Inst Sao Carlos, Trabalhador Sao Carlense Av 400, BR-13566590 Sao Carlos, SP - Brazil
[2] Univ Calif Santa Cruz, Dept Chem & Biochem, 1156 High St, Santa Cruz, CA 95060 - USA
[3] Univ Sao Paulo, Fac Sci Philosophy & Letters, BR-14040901 Ribeirao Preto, SP - Brazil
[4] State Univ Northern Rio de Janeiro Darcy Ribeiro, UENF, Phys Sci Lab, Av Alberto Lamego 2000, BR-28013602 Campos Dos Goytacazes, RJ - Brazil
[5] Univ British Columbia, Dept Biochem & Mol Biol, 2350 Hlth Sci Mall, Vancouver, BC V6T 1Z3 - Canada
[6] Sao Paulo State Univ, UNESP, Chem Inst, Dept Phys Chem, BR-14800900 Araraquara, SP - Brazil
[7] Sao Paulo State Univ, UNESP, Phys Dept, Inst Biosci Letters & Exact Sci IBILCE, Sao Jose Do Rio Preto, SP - Brazil
[8] Sao Paulo State Univ, UNESP, Inst Biosci Rio Claro, Dept Gen & Appl Biol, Rio Claro, SP - Brazil
[9] Univ Estadual Paulista, Biosci Inst, BR-18618000 Botucatu, SP - Brazil
[10] Univ Fed Sao Carlos, UFSCar, Dept Genet & Evolut, BR-13565905 Sao Carlos, SP - Brazil
Total Affiliations: 10
|
| Document type: | Journal article |
| Source: | Journal of Molecular Biology; v. 433, n. 23 NOV 19 2021. |
| Web of Science Citations: | 0 |
| Abstract | |
Several molecular mechanisms are involved in the genetic code interpretation during translation, as codon degeneration for the incorporation of rare amino acids. One mechanism that stands out is selenocysteine (Sec), which requires a specific biosynthesis and incorporation pathway. In Bacteria, the Sec biosynthesis pathway has unique features compared with the eukaryote pathway as Ser to Sec conversion mechanism is accomplished by a homodecameric enzyme (selenocysteine synthase, SelA) followed by the action of an elongation factor (SelB) responsible for delivering the mature Sec-tRNA(Sec) into the ribosome by the interaction with the Selenocysteine Insertion Sequence (SECIS). Besides this mechanism being already described, the sequential events for Sec-tRNA(Sec) and SECIS specific recognition remain unclear. In this study, we determined the order of events of the interactions between the proteins and RNAs involved in Sec incorporation. Dissociation constants between SelB and the native as well as unacylated-tRNA(Sec) variants demonstrated that the acceptor stem and variable arm are essential for SelB recognition. Moreover, our data support the sequence of molecular events where GTP-activated SelB strongly interacts with SelA. tRNA(Sec). Subsequently, SelB.GTP.tRNA(Sec) recognizes the mRNA SECIS to deliver the tRNA Sec to the ribosome. SelB in complex with its specific RNAs were examined using Hydrogen/Deuterium exchange mapping that allowed the determination of the molecular envelopes and its secondary structural variations during the complex assembly. Our results demonstrate the ordering of events in Sec incorporation and contribute to the full comprehension of the tRNA(Sec) role in the Sec amino acid biosynthesis, as well as extending the knowledge of synthetic biology and the expansion of the genetic code. (C) 2021 Elsevier Ltd. All rights reserved. (AU) | |
| FAPESP's process: | 13/17791-0 - Molecular and structural characterization of the Leishmania RNA virus LRV1-4 |
| Grantee: | Otavio Henrique Thiemann |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 14/00206-0 - Structure and function of SARS-CoV spike glycoprotein fusion peptides |
| Grantee: | Luís Guilherme Mansor Basso |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 12/15777-8 - Kinetic and structural studies of the Deoxycytidylate deaminase (DEOC) from Schistosoma mansoni |
| Grantee: | Jessica Fernandes Scortecci |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |
| FAPESP's process: | 12/23730-1 - Characterization of macromolecular interactions of proteins involved in the selenocysteine synthesis from Escherichia coli |
| Grantee: | Vitor Hugo Balasco Serrão |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 14/16005-4 - Characterization of the molecular interactions within selenocysteine synthesis pathway proteins |
| Grantee: | Jessica Fernandes Scortecci |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |
| FAPESP's process: | 98/14138-2 - Center for Structural Molecular Biotechnology |
| Grantee: | Glaucius Oliva |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |