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Microbiology. 2005 Dec;151(Pt 12):4033-4043. doi: 10.1099/mic.0.28219-0.

Probing the domain structure of FtsZ by random truncation and insertion of GFP.

Author information

1
Dept of Cell Biology, 3709 Duke University Medical Center, Durham, NC 27710, USA.

Abstract

Random transposon-mediated mutagenesis has been used to create truncations and insertions of green fluorescent protein (GFP), and Venus-yellow fluorescent protein (YFP), in Escherichia coli FtsZ. Sixteen unique insertions were obtained, and one of them, in the poorly conserved C-terminal spacer, was functional for cell division with the Venus-YFP insert. The insertion of enhanced GFP (eGFP) at this same site was not functional; Venus-YFP was found to be superior to eGFP in other respects too. Testing the constructs for dominant negative effects led to the following general conclusion. The N-terminal domain, aa 1-195, is an independently folding domain that can poison Z-ring function when expressed without a functional C-terminal domain. The effects were weak, requiring expression of the mutant at 3-5 times the level of wild-type FtsZ. The C-terminal domain, aa 195-383, was also independently folding, but had no activity in vivo. The differential activity of the N- and C-terminal domains suggests that FtsZ protofilament assembly is directional, with subunits adding primarily at the bottom of the protofilament. Directional assembly could occur by either a treadmilling or a dynamic instability mechanism.

PMID:
16339948
DOI:
10.1099/mic.0.28219-0
[Indexed for MEDLINE]

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