INTRODUCTION
Cellular division is a complex process required by all organisms for growth and differentiation. Cytokinesis and subsequent cell separation are either partially or completely dependent on the formation of an actomyosin contractile ring, depending on the particular organism. Actin and myosin are the major ring components and are supported by at least 130 other factors involved in cytokinesis (
42). Cytokinesis and cell separation in both fission and budding yeasts occur in an analogous manner, despite key differences between these organisms. In the budding yeast
Saccharomyces cerevisiae, the site of division is established either adjacent to or across from a previous division site during G
1 of the cell cycle (
30). The Rho GTPase Cdc42p is recruited to the presumptive site of division, where it is activated and recruits the septin scaffold proteins in late G
1 (
55). The sole type II myosin in
S. cerevisiae, encoded by
MYO1, is essential for actomyosin ring formation and is the first component of the ring to be assembled in a septin-dependent manner (
7,
30,
50). At the end of anaphase, F-actin forms a ring in association with Myo1p. The actomyosin ring contracts and results in the invagination of the plasma membrane at the neck between the mother and daughter cells. The actomyosin ring eventually disappears, and the cells are separated initially by a primary septum and then by flanking secondary septa made of chitin (
7). The cells separate after degradation of the primary septum, completing cytokinesis (
43).
In the fission yeast
Schizosaccharomyces pombe, the localization of the division site is selected before entry into mitosis (
27). Fission yeasts position the nascent septum at the cell midpoint using an interphase negative signal from a concentration gradient of the kinase Pom1p, which concentrates at both poles (
1,
33,
38), and the nucleus as a positional determinant, a process involving counterbalancing microtubule forces (
16,
51). Localization of Myo2p depends on phosphorylation in the C-terminal coiled coil and occurs in a septation initiation network (SIN)-dependent manner (
39). Cdc15p, the formin Cdc12p, ring assembly protein 2 Rng2p, and Mid1p simultaneously localize with Myo2p at the cytokinesis nodes (assemblies serving as the precursors of the contractile ring). The Rho GTPase Rho1p then regulates the formation of the ring through the activation of formins (
55). F-actin cables form at neighboring nodes, and Myo2p captures the actin filaments, applying force to bring the nodes together. By pulling the neighboring actin filaments, myosin proteins assemble and constrict the ring (
42). Unlike in
S. cerevisiae, the motor domain of
S. pombe type II myosins is required for cytokinesis (
31).
In contrast to yeast and animal cells, filamentous fungi undergo cytokinesis without cell separation during proliferative growth. This process is termed septation and results in the separation of hyphal cells by the formation of chitin-rich structures (septa) composed of several electron-dense layers that are perforated by a single pore (
36). This enables growth as multicellular compartmentalized hyphae. In
Aspergillus nidulans, the duplication cycle is initiated by waves of nuclear division that extend basally from the tips of apical cells, and this mitotic wave is followed by septum formation in the apical cell (
14,
26). Septation involves assembly of an actin ring between nuclei. Actin condensation and invagination initiation are closely followed by chitin synthesis at the edge of the invaginating actin ring. Actin withdraws from the developing septum in a punctuate form and eventually disappears, leaving the septal cell wall (
36). Similar to that in
S. cerevisiae and
S. pombe,
A. nidulans septum formation is actin dependent (
19). However, in contrast to the case in these yeasts, actin localizes simultaneously at the tips of growing cells and at the site of septum formation (
10,
19). In addition, microtubules are also required for the initiation and progression of septation (
36).
Penicillium marneffei is a thermally dimorphic fungal pathogen which uses three different modes of division during the various stages of its life cycle (
3). At 25°C, it grows in a multinucleate, branched, septate hyphal form by apical extension in a mode similar to that of most other filamentous fungi. Under the appropriate environmental conditions, hyphal cells undergo asexual development to produce conidiophores. The differentiated cell types present in the conidiophore emerge from a stalk cell by a sequential budding process which requires coupling of nuclear division and cell division. Cell separation is required to liberate the uninucleate asexual spores (conidia) from the terminal end of the differentiated conidiophore. At 37°C,
P. marneffei undergoes a process termed arthroconidiation, where cellular division and nuclear division become coupled, hyphae lay down double septa, and cells subsequently separate to liberate uninucleate yeast cells. The yeast cells proliferate vegetatively by fission division. The capacity for three modes of cellular division in a single organism provides a unique system in which to probe the similarities and differences between these processes. Previous studies with
P. marneffei have shown specialization in the control of these different modes of division by three small GTPase-encoding genes, with concomitant overlapping roles during cytokinesis (
9–11). Therefore,
P. marneffei is an excellent organism in which to compare the differences and similarities between the three modes of cellular division exhibited by fungi and whether the formation of an actomyosin ring is required for completion of cytokinesis in the different modes of division. Here we describe the cloning and characterization of a gene (
myoB) encoding a type II myosin from
P. marneffei and investigate its role in cytokinesis during the three modes of cellular division (hyphal growth, conidiation, and yeast morphogenesis).