The Candida boidinii peroxisomal membrane protein Pmp30 has a role in peroxisomal proliferation and is functionally homologous to Pmp27 from Saccharomyces cerevisiae

Abstract

The mechanism of peroxisome proliferation is poorly understood. Candida boidinii is a methylotrophic yeast that undergoes rapid and massive peroxisome proliferation and serves as a good model system for this process. Pmp30A and Pmp30B (formerly designated Pmp31 and Pmp32, respectively) are two closely related proteins in a polyploid strain of this yeast that are strongly induced by diverse peroxisome proliferators such as methanol, oleate, and D-alanine. The function of these proteins is not understood. To study this issue, we used a recently described haploid strain (S2) of C. boidinii that can be manipulated genetically. We now report that strain S2 contains a single PMP30 gene very similar in sequence (greater than 93% identity at the DNA level) to PMP30A and PMP30B. When PMP30 was disrupted, cell growth on methanol was greatly inhibited, and cells grown in both methanol and oleate had fewer, larger, and more spherical peroxisomes than wild-type cells. A similar phenotype was recently described for Saccharomyces cerevisiae cultured on oleate in which PMP27, which encodes a protein of related sequence that is important for peroxisome proliferation, was disrupted. To determine whether Pmp27 is a functional homolog of Pmp30, genetic complementation was performed. PMP30A was expressed in the PMP27 disruptant of S. cerevisiae, and PMP27 was expressed in the PMP30 disruptant of C. boidinii S2. Complementation, in terms of both cell growth and organelle size, shape, and number, was successful in both directions, although reversion to a wild-type phenotype was only partial for the PMP30 disruptant. We conclude that these proteins are functional homologs and that both Pmp30 and Pmp27 have a direct role in proliferation and organelle size rather than a role in a specific peroxisomal metabolic pathway of substrate utilization.

Pex30p, Pex31p, and Pex32p Form a Family of Peroxisomal Integral Membrane Proteins Regulating Peroxisome Size and Number inSaccharomyces cerevisiae

“…In normal human cells, in contrast, conversion of immature peroxisomal vesicles to mature peroxisomes by membrane and matrix protein import may occur before peroxisomes undergo division (Gould and Valle, 2000). Members of the Pex11 family of peroxins, including Pex25p and Pex27p Rottensteiner et al, 2003b) of S. cerevisiae, have been shown to effect peroxisome division in different organisms (Erdmann and Blobel, 1995;Marshall et al, 1995;Sakai et al, 1995;. The dynamin-like protein Vps1p has also been implicated in this process (Hoepfner et al, 2001), and we recently showed that the peroxisomal integral membrane proteins Pex28p and Pex29p are also involved in controlling the number, size, and separation of peroxisomes in S. cerevisiae .…”

“…Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc. E03-09 -0681.peroxisomes, whereas cells lacking Pex11p have fewer but larger peroxisomes than normal (Erdmann and Blobel, 1995;Marshall et al, 1995;Sakai et al, 1995;. The dynamin-like protein Vps1p (Hoepfner et al, 2001) and the peroxins Pex25p and Pex27p Rottensteiner et al, 2003b) of the yeast Saccharomyces cerevisiae have recently been shown to be required for the control of peroxisome size and number.…”

Clofibrate‐inducible, 28‐kDa peroxisomal integral membrane protein is encoded by PEX11

“…We herein isolated human PEX11 cDNA (HsPEX11) encoding a peroxin Pex11p, by EST homology search using Candida boidinii (Cb) PEX11 [36]. Human Pex11p was recognized by antibody raised against clo¢brate-inducible, 28-kDa peroxisomal membrane protein (PMP28).…”

The Dynamin-like GTPase DLP1 Is Essential for Peroxisome Division and Is Recruited to Peroxisomes in Part by PEX11

“…Of the many PEX 1 genes and products (peroxins) required for peroxisome biogenesis, only PEX11 has been shown to have a conserved role in peroxisome division (7)(8)(9)(10)(11)(12)(13). The overexpression of PEX11 promotes peroxisome elongation and subsequent division, whereas loss of PEX11 results in reduced peroxisome abundance (7,8,12,14).…”