25º Congresso Brasileiro de Microbiologia
ResumoID:2120-1


Área: Genética e Biologia Molecular ( Divisão N )

ANALYSIS OF THE CELL DIVISION COMPLEX OF BACILLUS SUBTILIS USING GFP FLUORESCENCE MUCROSCOPY਀㰀⼀猀琀爀漀渀最㸀㰀⼀瀀㸀㰀瀀 愀氀椀最渀㴀樀甀猀琀椀昀礀㸀㰀戀㸀㰀甀㸀䜀甀椀氀栀攀爀洀攀 䰀漀甀稀愀搀愀 匀椀氀瘀愀 䴀攀椀爀愀 㰀⼀甀㸀㰀⼀戀㸀  ⠀㰀椀㸀唀匀倀㰀⼀椀㸀⤀㬀 㰀戀㸀䘀爀攀搀攀爀椀挀漀 䨀漀猀攀  䜀甀攀椀爀漀猀ⴀ昀椀氀栀漀 㰀⼀戀㸀  ⠀㰀椀㸀唀匀倀㰀⼀椀㸀⤀㰀戀爀㸀㰀戀爀㸀㰀⼀瀀㸀㰀戀㸀㰀昀漀渀琀 猀椀稀攀㴀㈀㸀刀攀猀甀洀漀㰀⼀昀漀渀琀㸀㰀⼀戀㸀㰀瀀 愀氀椀最渀㴀樀甀猀琀椀昀礀 挀氀愀猀猀㴀琀爀攀猀㸀㰀昀漀渀琀 猀椀稀攀㴀㈀㸀

Bacillus subtilis undergoes binary fission, with the septum forming in the middle of a progenitor cell, resulting in genetically identical daughter cells. Septum formation in bacteria is carried out by a large protein complex known as the divisome. FtsZ (a tubulin homolog) self-assembles in a structure known as "Z ring" in the middle of the cell. The Z ring is the first event in divisome formation and it is responsible to recruit other division proteins to the divisome. The proteins that constitute the divisome complex are well known in the literature, but how the divisome is assembled remains unclear. In this work we have used fluorescence microscopy as a tool to study the kinetics of divisome formation. We created a red fluorescent FtsZ fusion (FtsZ-mCherry) and used it to measure the frequency of co-localization between the Z ring and five divisome proteins (ZapA, EzrA, DivIVA, YpsB, MinC), each individually fused to GFP. By measuring the frequency of co-localization between the Z ring and other division proteins we can establish the sequence of events during divisome assembly. We have found that ZapA, EzrA, YpsB, DivIVA and MinC co-localized with the Z-ring in 97%, 98%, 50%, 42%, 32% of the cells, respectively. The high frequency of co-localization with FtsZ suggests that ZapA and EzrA like to assemble into the divisome immediately after Z ring formation. On the other hand, YpsB, DivIVA and MinC, co-localize much less with the Z ring and, thus, seem to exhibit a delay in their association with the complex. The different co-localization frequencies among proteins suggest that the divisome does not assemble instantly. Instead, it seems to mature in a more gradual way, with each protein arriving at the complex with a different kinetics.

 

 

Financial Support: Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP)

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