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Nat Commun ; 15(1): 4850, 2024 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-38844782

RESUMO

Bacterial RNAP needs to form holoenzyme with σ factors to initiate transcription. While Staphylococcus aureus σA controls housekeeping functions, S. aureus σB regulates virulence, biofilm formation, persistence, cell internalization, membrane transport, and antimicrobial resistance. Besides the sequence difference, the spacers between the -35 element and -10 element of σB regulated promoters are shorter than those of σA regulated promoters. Therefore, how σB recognizes and initiates transcription from target promoters can not be inferred from that of the well studied σ. Here, we report the cryo-EM structures of S. aureus RNAP-promoter open complexes comprising σA and σB, respectively. Structural analyses, in combination with biochemical experiments, reveal the structural basis for the promoter specificity of S. aureus transcription. Although the -10 element of σA regulated promoters is recognized by domain σA2 as single-stranded DNA, the -10 element of σB regulated promoters is co-recognized by domains σB2 and σB3 as double-stranded DNA, accounting for the short spacers of σB regulated promoters. S. aureus RNAP is a validated target of antibiotics, and our structures pave the way for rational drug design targeting S. aureus RNAP.


Assuntos
Proteínas de Bactérias , Microscopia Crioeletrônica , RNA Polimerases Dirigidas por DNA , Regiões Promotoras Genéticas , Fator sigma , Staphylococcus aureus , Staphylococcus aureus/genética , Staphylococcus aureus/enzimologia , RNA Polimerases Dirigidas por DNA/metabolismo , RNA Polimerases Dirigidas por DNA/genética , RNA Polimerases Dirigidas por DNA/química , Fator sigma/metabolismo , Fator sigma/genética , Fator sigma/química , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/química , Regulação Bacteriana da Expressão Gênica , Modelos Moleculares , Transcrição Gênica , Ligação Proteica
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