Project name: Metamycoplasma hominis

Description: For a comprehensive characterisation of the M. hominis-action in infection a customized Mho microarray, which was based on two genome sequences (PG21 and LBD-4), was newly designed and validated for M. hominis strain FBG, to analyze the dynamics of the mycoplasma transcriptome during infection. RNA-preparation was evaluated and adopted for highest recovery of mycoplasmal mRNAs from in vitro HeLa cell infection assays. Following cRNA hybridization, the read out strategy of the hybridization results was optimized and confirmed by RT-PCR. A statistically robust infection assay with M. hominis strain FBG enabled the identification of regulated key effector molecules such as critical cytoadhesins (4 h post infection (pI)), invasins (48h pI) and proteins associated with establishing chronic infection of the host (336h pI). Of the 304 differentially regulated genes 130 (43%) encoded hypothetical proteins, including lipoproteins that seem to play a central role as virulence factors at each stage of infection: P75 as novel cytoadhesin candidate, which is also upregulated in the chronic infection, the MHO_2100-protein as postulated invasin and the MHO_730-protein as novel ecto-nuclease and domain of a novel ABC transporter, whose function in chronic infection has to be elucidated in the future. Implementation of the M. hominis microarray strategy led to a comprehensive identification of to date unknown candidates for virulence-factors at relevant stages of host cell infection.Overall design: A customized Mho microarray was designed with 13,708 non redundant Mho oligos in total, validated and used for hybridization of cRNA of total RNA from HeLa infection assays, which were prepared in duplicates after 1h, 4h, 48h and 336 h post infection with 50 MOI of M. hominis strain FBG. RNA of time point 1h of infection was used as a reference.

Data type: Transcriptome or Gene expression

Sample scope: Multiisolate

Relevance: Other

Last updated: 2017-04-10