Fig. 2: Identification of PrfaH target sites and structural analysis.

a The deletion of the intergenic region (IGR) between rs000240 and virB1 does not affect plasmid conjugation. The diagram shows mutations in IGR. ∆IGR refers to the pHNYX644∆IGR plasmid. A synthetic SD sequence (5’-AAAGAGGAGAAA-3’) was fused upstream of virB1, resulting in the pHNYX644∆IGR-SD plasmid. b The 5’UTR of PrfaH is indispensable for the function of PrfaH. The diagram shows mutations in the upstream region of prfaH. ∆prfaH refers to the pHNYX644∆prfaH plasmid. ∆prfaH-PJ23119 represents a construct where the prfaH promoter was replaced with the PJ23119 promoter in the pHNYX644∆prfaH plasmid, with a synthetic SD sequence fused upstream of prfaH. ∆prfaH-PJ23119-UTR indicates that the synthetic SD sequence was replaced by the 5’ UTR of prfaH. Dashed arrows without color annotations indicate gene deletions. c Identification of the target of PrfaH in 5’UTR of PrfaH. The pHNYX644∆prfaH plasmids with mutations in the 5’UTR of prfaH (W1 to W12), and the mutated bases are indicated in red font. ∆prfaH represents the pHNYX644∆prfaH plasmid. The conjugation rate of pHNYX644∆prfaH and its derivatives with mutations in the 5’UTR of PrfaH were measured in the presence or absence of PrfaH. The conjugation rates of all prfaH-complemented mutants were compared with the conjugation rate of ∆prfaH complemented with prfaH using t-test with Welch’s correction. Conjugation assays were conducted with n = 3 biological replicates, and statistical analysis was performed using t-test with Welch’s correction.