Identification of novel components of the Ced and Ups systems in Saccharolobus islandicus REY15A.

Abstract

In Sulfolobales cells, transcription of the Ups (UV-inducible pili of Sulfolobus) and Ced (Crenarchaeal system for exchange of DNA) genes is highly induced by DNA damage, and the two systems play key roles in pili-mediated cell aggregation and chromosomal DNA import, respectively. Ups is composed of UpsA, UpsB, UpsE, and UpsF, while Ced is composed of CedA, CedA1, CedA2, and CedB. So far, how DNA is transported by these systems is far from clear. Here, we report three novel components of the Ced and Ups systems in Saccharolobus islandicus REY15A, CedD (SiRe_1715) and CedE (SiRe_2100), paralogs of CedB and CedA, and UpsC (SiRe_1957), a paralog of UpsA/UpsB. We developed a DNA import and export assay method, by which we revealed that CedD, CedE, and UpsC are essential for DNA import, while CedE and UpsC are also involved in DNA export together with CedA1 and Ups. Microscopic analysis revealed that upsC is involved in cell aggregation like other Ups genes. In addition, we found that cedB and cedD co-occur in the Crenarchaeal genomes that lack virB4, an essential component of type IV secretion system. Interestingly, CedB and CedD share homology to different parts of VirB4 N-terminal domain and form stable homo-oligomers in vitro. Collectively, our results indicate that CedD, CedE, and UpsC are integral components of the Ced and Ups systems in Sulfolobales.