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Volume 151, Issue 12

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

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 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 . 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.

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  • Published Online:
Keyword(s): eGFP, enhanced green fluorescent protein , GFP, green fluorescent protein and YFP, yellow fluorescent protein
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2005-12-01
2024-04-20
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Probing the domain structure of FtsZ by random truncation and insertion of GFP
Microbiology 151, 4033 (2005); https://doi.org/10.1099/mic.0.28219-0
/content/journal/micro/10.1099/mic.0.28219-0
/content/journal/micro/10.1099/mic.0.28219-0
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