Septin

Available protein structures:
Pfam	structures / ECOD
PDB	RCSB PDB; PDBe; PDBj
PDBsum	structure summary

Cell division/GTP binding protein
Cell division/GTP binding protein
Identifiers
Symbol	Cell_Div_GTP_bd
Pfam	PF00735
Pfam clan	CL0023
InterPro	IPR000038
Pfam
Available protein structures:
Pfam	structures / ECOD
PDB	RCSB PDB; PDBe; PDBj
PDBsum	structure summary

Septins are evolutionary conserved proteins with essential functions in cytokinesis, and more subtle roles throughout the cell cycle. Much of our knowledge about septins originates from studies of budding yeast Saccharomyces cerevisiae, where they form a ring-like protein scaffold at the mother-bud neck. Recent research in human cells suggests that septins build 'cages' around bacterial pathogens, immobilizing the harmful microbes and preventing them from invading other cells.^[1]

Septins in Saccharomyces cerevisiae

History

The septins were discovered in 1970 by Leland H. Hartwell and colleagues in a screen for temperature-sensitive mutants affecting cell division (cdc mutants). The screen revealed four mutants which prevented cytokinesis at restrictive temperature. The corresponding genes represent the four original septins, ScCDC3, ScCDC10, ScCDC11, and ScCDC12. Despite disrupted cytokinesis, the cells continued budding, DNA synthesis, and nuclear division, which resulted in large multinucleate cells with multiple, elongated buds. In 1976, analysis of electron micrographs revealed ~20 evenly spaced striations of 10-nm filaments around the mother-bud neck in wild-type but not in septin-mutant cells. Immunofluorescence studies revealed that the septin proteins colocalize into a septin ring at the neck. The localization of all four septins is disrupted in conditional Sccdc3 and Sccdc12 mutants, indicating interdependence of the septin proteins. Strong support for this finding was provided by biochemical studies: The four original septins co-purified on affinity columns, together with a fifth septin protein, encoded by ScSEP7 or ScSHS1. Purified septins from budding yeast, Drosophila, Xenopus, and mammalian cells are able to self associate in vitro to form highly ordered, filamentous structures. How the septins interact in vitro to form heteropentamers that assemble into filaments was studied in detail in S. cerevisiae. Based on these and former studies, the septins are composed of a variable N-terminus with a basic phosphoinositide binding motif, a conserved core comprising a GTP-binding domain, a septin-unique element and a C-terminal extension including a predicted coiled coil.

Micrographs of purified filaments raised the possibility that the septins are organized in parallel to the mother-bud axis. The 10-nm striations seen on electron micrographs may be the result of lateral interaction between the filaments. Mutant strains lacking factors important for septin organization support this view. Instead of continuous rings, the septins form bars oriented along the mother-bud axis in deletion mutants of ScGIN4, ScNAP1 and ScCLA4.

Functions

Scaffold

The septins act as a scaffold, recruiting a plethora of proteins. These protein complexes are involved in cytokinesis, chitin deposition, cell polarity, spore formation, in the morphogenesis checkpoint, spindle alignment checkpoint and bud site selection.

Cytokinesis

Budding yeast cytokinesis is driven through two septin dependent, redundant processes: recruitment and contraction of the actomyosin ring and formation of the septum by vesicle fusion with the plasma membrane. In contrast to septin mutants, disruption of one single pathway only leads to a delay in cytokinesis, not complete failure of cell division. Hence, the septins are predicted to act at the most upstream level of cytokinesis.

Cell polarity

After the apical-isotropic switch in budding yeast, cortical components, supposedly of the exocyst and polarisome, are delocalized from the apical pole to the entire plasma membrane of the bud, but not the mother cell. The septin ring at the neck serves as a cortical barrier that prevents membrane diffusion of these factors between the two compartments. This asymmetric distribution is abolished in septin mutants.

Some conditional septin mutants do not form buds at their normal axial location. Moreover, the typical localization of some bud-site-selection factors in a double ring at the neck is lost or disturbed in these mutants. This indicates that the septins may serve as anchoring site for such factors in axially budding cells.

Organization

It seems that one single septin organization should not be sufficient to fulfill such a variety of tasks. Accordingly, the septin cortex undergoes several changes throughout the cell cycle: The first visible septin structure is a distinct ring which appears ~15 min before bud emergence. After bud emergence, the ring broadens to assume the shape of an hourglass around the mother-bud neck. During cytokinesis, the septin cortex splits into a double ring which eventually disappears. How can the septin cortex undergo so dramatic changes, although some of its functions may require it to be a stable structure? FRAP analysis has revealed that the turnover of septins at the neck undergoes multiple changes during the cell cycle. The predominant, functional conformation is characterized by a low turnover rate (frozen state), during which the septins are phosphorylated. Structural changes require a destabilization of the septin cortex (fluid state) induced by dephosphorylation prior to bud emergence, ring splitting and cell separation.

The composition of the septin cortex does not only vary throughout the cell cycle but also along the mother-bud axis. This inherent polarity of septin filaments allows concentration of some proteins primarily to the mother side of the neck, some to the center and others to the bud site.

Septins in filamentous fungi

Since their discovery in S. cerevisiae, septin homologues have been found throughout the eukaryotic kingdom, with the exception of plants. The variety of different shapes that septins can assume within a single cell is especially apparent in filamentous fungi, where they control aspects of filamentous morphology.

Candida albicans

The genome of C. albicans encodes homologues to all S. cerevisiae septins (CaCDC3, CaCDC10, CaCDC11, CaCDC12, CaSEP7). They form a diffuse band at the base of emerging hyphae, a bright double ring at septation sites, an extended diffuse cap at hyphal tips and elongated filaments stretching around the spherical chlamydospores. As an effect of maturation, double rings reflect hyphal polarity by disassembling the tip proximal ring. CaCdc3p and CaCdc12p are essential for proliferation in yeast or hyphal forms. Cacdc10Δ and Cacdc11Δ deletion mutants are viable but show aberrant chitin localization and cannot properly maintain hyphal growth direction.

Aspergillus nidulans

Five septins are found in A. nidulans (AnAspAp, AnAspBp, AnAspCp, AnAspDp, AnAspEp). AnAspBp forms single rings at septation sites that eventually split into double rings. Additionally, AnAspBp forms a ring at sites of branch emergence which broadens into a band as the branch grows. Like in C. albicans, double rings reflect polarity of the hypha, but by disassembling the more basal ring. Bases for the various patterns of septin organization could be different modifications and/or localization of different septin interaction partners. Conditional mutants of the essential AnAspBp display diffuse chitin deposition and a hyper-branching phenotype.

Ashbya gossypii

The ascomycete A. gossypii possesses homologues to all S. cerevisiae septins, with one being duplicated (AgCDC3, AgCDC10, AgCDC11A, AgCDC11B, AgCDC12, AgSEP7). In vivo studies of AgSep7p-GFP have revealed that septins assemble into discontinuous hyphal rings close to growing tips and sites of branch formation and into asymmetric structures at the base of branching points. Rings are made of filaments which are long and diffuse close to growing tips and short and compact further away from the tip. During septum formation, the septin ring splits into two to form a double ring. Agcdc3Δ, Agcdc10Δ and Agcdc12Δ deletion mutants display aberrant morphology and are defective for actin-ring formation, chitin-ring formation, and sporulation. Due to the lack of septa, septin deletion mutants are highly sensitive, and damage of a single hypha can result into complete lysis of a young mycelium.

Human septins

However, most studies of septins, or guanosine-5′-triphosphate (GTP) binding proteins, have been confined to yeast cells. The latest research in human cells suggests that septins build 'cages' around bacterial pathogens, immobilizing the harmful microbes and preventing them from invading other healthy cells. This cellular defence system could be explored to create therapies for dysentery and other illnesses. “This is a new way for cells to control an infection,” Shigella, a bacterium that causes sometimes lethal diarrhoea in humans and other primates. To propagate from cell to cell, Shigella bacteria develop actin-polymer 'tails', which propel the microbes around and allow them to force their way into neighbouring host cells. To counterattack, human cells produce a cell-signalling protein called TNF-α. The researchers found that when TNF-α is present, thick bundles of septin filaments encircle the microbes. This, in turn, interferes with tail formation and stops Shigella in its tracks. Microbes that become trapped in septin cages are broken down in a stage of the cell's life cycle called autophagy. “Autophagy is more efficient because of the septin cage, and the septin cage does not occur if you do not have the autophagy. Many research groups are working on understand the link between septins and autophagy, and to determine how important septins are in humans in vivo. Disruptions in septins and mutations in the genes that code for them could be involved in causing leukaemia, colon cancer and neurodegenerative conditions such as Parkinson’s disease and Alzheimer’s disease. Potential therapies for these, as well as for bacterial conditions such as dysentery caused by Shigella, might bolster the body’s immune system with drugs that mimic the behaviour of TNF-α and allow the septin cages to proliferate.^[1]

In mitochondria

The septin localized in the mitochondria is called mitochondrial septin (M-septin). It was identified as a CRMP/CRAM-interacting protein in developing mouse brain.^[2]

References

^ ^a ^b Mascarelli A (2011). "Septin proteins take bacterial prisoners: A cellular defence against microbial pathogens holds therapeutic potential". Nature. doi:10.1038/nature.2011.9540. {{cite journal}}: Unknown parameter |month= ignored (help)
^ Takahashi S, Inatome R, Yamamura H, Yanagi S (2003). "Isolation and expression of a novel mitochondrial septin that interacts with CRMP/CRAM in the developing neurones". Genes Cells. 8 (2): 81–93. doi:10.1046/j.1365-2443.2003.00617.x. PMID 12581152. {{cite journal}}: Unknown parameter |month= ignored (help)CS1 maint: multiple names: authors list (link)