FtsZ Protofilament Curvature is the Opposite of Tubulin Rings

Bacterial tubulin homolog FtsZ assembles straight protofilaments (pfs) that form the scaffold of the cytokinetic Z ring. These pfs can adopt a curved conformation forming a miniring or spiral tube 24 nm in diameter. Tubulin pfs also have a curved conformation, forming 42 nm tubulin rings. We have previously provided evidence that FtsZ generates a constriction force by switching from straight pfs to the curved conformation, generating a bending force on the membrane. In the simplest model the membrane tether, which exits from the C terminus of the globular FtsZ, would have to be on the outside of the curved pf. However, it is well established that tubulin rings have the C terminus on the inside of the ring. Could FtsZ and tubulin rings have the opposite curvature? In the present study we explored the direction of curvature of FtsZ rings by fusing large protein tags to the N or C terminus of the FtsZ globular domain. FtsZ with a protein tag on the N terminus did not assemble tubes. This was expected if the N terminus is on the inside, because the protein tags are too big to fit in the interior of the tube. FtsZ with C-terminal tags assembled normal tubes, consistent with the C terminus on the outside. The FN extension was not visible in negative stain, but thin section EM gave definitive evidence that the C-terminal tag was on the outside of the tubes. This has interesting implications for the evolution of tubulin. It seems likely that tubulin began with the curvature of FtsZ, which would have resulted in pfs curving toward the interior of a disassembling MT. Evolution not only eliminated this undesirable curvature, but managed to reverse direction to produce the outward curving rings, which is useful for pulling chromosomes.

Structural manifestation of the delocalization error of density functional approximations: C(4N+2) rings and C(20) bowl, cage, and ring isomers.

The ground state structure of C(4N+2) rings is believed to exhibit a geometric transition from angle alternation (N < or = 2) to bond alternation (N > 2). All previous density functional theory (DFT) studies on these molecules have failed to reproduce this behavior by predicting either that the transition occurs at too large a ring size, or that the transition leads to a higher symmetry cumulene. Employing the recently proposed perspective of delocalization error within DFT we rationalize this failure of common density functional approximations (DFAs) and present calculations with the rCAM-B3LYP exchange-correlation functional that show an angle-to-bond-alternation transition between C(10) and C(14). The behavior exemplified here manifests itself more generally as the well known tendency of DFAs to bias toward delocalized electron distributions as favored by Huckel aromaticity, of which the C(4N+2) rings provide a quintessential example. Additional examples are the relative energies of the C(20) bowl, cage, and ring isomers; we show that the results from functionals with minimal delocalization error are in good agreement with CCSD(T) results, in contrast to other commonly used DFAs. An unbiased DFT treatment of electron delocalization is a key for reliable prediction of relative stability and hence the structures of complex molecules where many structure stabilization mechanisms exist.