![]() aeruginosa, providing a structural basis of how the CupE1 protein forms zigzag-shaped, extended pili. In this study, we present a cryo-EM structure of the archaic CUP pilus CupE from P. Archaic CUP pili help maintain the architecture of multicellular bacterial aggregates termed biofilms formed by the pathogen Pseudomonas aeruginosa and many others. However, little is known about the so-called archaic CUP pili, which are the most widespread type. Previous studies have characterised various Chaperone-Usher Pathway (CUP) pili, which are common in Gram-negative bacteria. Many bacteria adhere to surfaces or host cells using filamentous structures termed pili that extend from the bacterial cell and anchor them to their target. Taken together, our study provides insights into the architecture of archaic CUP pili, providing a structural basis for understanding their role in promoting cellular adhesion and biofilm formation in P. aeruginosa and the co-occurrence of cupE with other cup clusters, suggesting interdependence of cup pili in regulating bacterial adherence within biofilms. Finally, bioinformatic analysis shows the widespread abundance of cupE genes in isolates of P. aeruginosa cells using electron cryotomography shows that CupE pili adopt variable curvatures in response to their environment, which might facilitate their role in promoting cellular attachment. ![]() We show that CupE1 subunits within the pilus are arranged in a zigzag architecture, containing an N-terminal donor β-strand extending from each subunit into the next, where it is anchored by hydrophobic interactions, with comparatively weaker interactions at the rest of the inter-subunit interface. In this study, we present the electron cryomicroscopy structure of the archaic CupE pilus from the opportunistic human pathogen Pseudomonas aeruginosa. While classical CUP pili have been extensively characterized, little is known about so-called archaic CUP pili, which are phylogenetically widespread and promote biofilm formation by several human pathogens. Chaperone-Usher Pathway (CUP) pili are major adhesins in Gram-negative bacteria, mediating bacterial adherence to biotic and abiotic surfaces. ![]()
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