Extensive diversity in RNA termination and regulation revealed by transcriptome mapping for the Lyme pathogenB. burgdorferi

Abstract

Transcription termination is an essential and dynamic process that can tune gene expression in response to diverse molecular signals. Yet, the genomic positions, molecular mechanisms, and regulatory consequences of termination have only been studied thoroughly in model bacteria. We employed complementary RNA-seq approaches to map RNA ends for the transcriptome of the spirochete Borrelia burgdorferi, the etiological agent of Lyme disease. By systematically mapping B. burgdorferi RNA ends at single nucleotide resolution, we delineated complex gene arrangements and operons and mapped untranslated regions (UTRs) and small RNAs (sRNAs). We experimentally tested modes of B. burgdorferi transcription termination and compared our findings to observations in E. coli, P. aeruginosa, and B. subtilis. We discovered 63% of B. burgdorferi RNA 3′ ends map upstream or internal to open reading frames (ORFs), suggesting novel mechanisms of regulation. Northern analysis confirmed the presence of stable 5′ derived RNAs from mRNAs encoding gene products involved in the unique infectious cycle of B. burgdorferi. We suggest these RNAs resulted from premature termination and regulatory events, including forms of cis-acting regulation. For example, we documented that the polyamine spermidine globally influences the generation of truncated mRNAs. In one case, we showed that high spermidine concentrations increased levels of RNA fragments derived from an mRNA encoding a spermidine import system, with a concomitant decrease in levels of the full-length mRNA. Collectively, our findings revealed new insight into transcription termination and uncovered an abundance of potential RNA regulators.

Broadly conserved FlgV controls flagellar assembly andBorrelia burgdorferidissemination in mice

“…The gene is embedded within the “ flgB operon” and expression of the entire operon has been suggested to be constitutive, driven by a consensus σ 70 promoter upstream of flgB (Ge et al ., 1997b, Zhang et al ., 2020). However, B. burgdorferi transcriptome mapping studies (Adams et al ., 2017, Petroni et al ., 2023) documented three predominant transcription start sites (TSSs), several 5′ processed ends, and several 3′ ends throughout the operon (Figure S5A). Therefore, this genomic region likely encodes multiple RNA products.…”