Novel Role of Phosphorylation-Dependent Interaction between FtsZ and FipA in Mycobacterial Cell Division
Figure 1
FipA regulates growth and cell division.
A. Growth of M. smegmatis mc2 155 (WT), FipA-KO (KO) or FipA-KO complemented with fipAMTB (WT) or its fipAMTB(T77A) [complemented (T77A)], was followed by monitoring cfu at different periods of time. B. Cultures were treated with H2O2 as described under “Materials and Methods.” Growth was monitored by determining cfu. Percent survival was determined with respect to the cfu of untreated cells (100%). C. Cell length measurements of wild type M. smegmatis (WT), or FipA-KO (KO), or FipA-KO complemented with fipAMTB (complemented) after treatment with H2O2 as described under “Materials and Methods.” At least 100 cells from each set were measured. D. Western blotting of lysates of M. tuberculosis expressing an antisense construct of fipA under the control of the hsp60 promoter, with anti-FipA antibody. The blot was reprobed with anti-FtsW as loading control. E. THP1 cells were infected with M. tuberculosis harbouring vector alone or an antisense construct of fipA at an MOI of 2 for 90 min; washed with fresh medium and grown for different periods of time. Internalized bacteria were released by lysing the macrophages followed by determination of cfu. M. tuberculosis harbouring vector alone, -▵-; or antisense construct of fipA, -•-. F, G. THP-1 cells were infected and intracellular bacteria were obtained by lysing the macrophages, as in (E). Cell length measurements were done for intracellular M. tuberculosis harbouring vector alone (F) or expressing an antisense construct of fipA under the control of the hsp60 promoter (G) after 1, 3, 5, or 6 days of growth in THP1 cells. At least 100 cells from each set were measured.