Transport of microinjected alcohol oxidase from Pichia pastoris into vesicles in mammalian cells: involvement of the peroxisomal targeting signal.

Walton, Paul A.; Gould, Stephen J.; Rachubinski, Richard A.; Subramani, Suresh; Feramisco, James R.

doi:10.1083/jcb.118.3.499

Cited by 31 publications

(18 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, in Y. lipolytica, the secretory pathway can play an essential role in peroxisome biogenesis. A role for the ER in the trafficking of membrane proteins to peroxisomes in mammalian and yeast cells has also been suggested by some recent observations (2,7,17,60).…”

Section: Discussionmentioning

confidence: 81%

Four Distinct Secretory Pathways Serve Protein Secretion, Cell Surface Growth, and Peroxisome Biogenesis in the Yeast Yarrowia lipolytica

Titorenko

Ogrydziak

Rachubinski

1997

Molecular and Cellular Biology

Self Cite

116

View full text Add to dashboard Cite

We have identified and characterized mutants of the yeast Yarrowia lipolytica that are deficient in protein secretion, in the ability to undergo dimorphic transition from the yeast to the mycelial form, and in peroxisome biogenesis. Mutations in the SEC238, SRP54, PEX1, PEX2, PEX6, and PEX9 genes affect protein secretion, prevent the exit of the precursor form of alkaline extracellular protease from the endoplasmic reticulum, and compromise peroxisome biogenesis. The mutants sec238A, srp54KO, pex2KO, pex6KO, and pex9KO are also deficient in the dimorphic transition from the yeast to the mycelial form and are affected in the export of only plasma membrane and cell wall-associated proteins specific for the mycelial form. Mutations in the SEC238, SRP54, PEX1, and PEX6 genes prevent or significantly delay the exit of two peroxisomal membrane proteins, Pex2p and Pex16p, from the endoplasmic reticulum en route to the peroxisomal membrane. Mutations in the PEX5, PEX16, and PEX17 genes, which have previously been shown to be essential for peroxisome biogenesis, affect the export of plasma membrane and cell wall-associated proteins specific for the mycelial form but do not impair exit from the endoplasmic reticulum of either Pex2p and Pex16p or of proteins destined for secretion. Biochemical analyses of these mutants provide evidence for the existence of four distinct secretory pathways that serve to deliver proteins for secretion, plasma membrane and cell wall synthesis during yeast and mycelial modes of growth, and peroxisome biogenesis. At least two of these secretory pathways, which are involved in the export of proteins to the external medium and in the delivery of proteins for assembly of the peroxisomal membrane, diverge at the level of the endoplasmic reticulum.The secretory pathway of eukaryotic cells consists of a series of morphologically and biochemically distinct membranebound compartments. The classical secretory pathway starts with protein translocation into the lumen of the endoplasmic reticulum (ER). From the ER, secretory proteins are transported within a series of vesicles to and through the Golgi complex and are then either delivered to the cell surface or routed to the endosomal-lysosomal (vacuolar) branch of the pathway (32,43,53). At all stages of the pathway, intercompartmental transport is initiated by the formation of coated vesicles on the donor compartment, followed by uncoating of vesicle intermediates prior to their fusion with a specific acceptor compartment (47,(52)(53)(54). While initial studies suggested the existence of one major pathway of vesicle-mediated protein export to the cell surface through a series of membrane-bound compartments, exceptions to this classical scheme of protein secretion have now been described.First, not all proteins of either mammalian or yeast cells are delivered to the cell surface via the vesicle-mediated secretory pathway. At least two nonclassical secretory pathways have been demonstrated in the yeast Saccharomyces cerevisiae (for a detailed discussion, s...

show abstract

Section: Discussionmentioning

confidence: 81%

Four Distinct Secretory Pathways Serve Protein Secretion, Cell Surface Growth, and Peroxisome Biogenesis in the Yeast Yarrowia lipolytica

Titorenko

Ogrydziak

Rachubinski

1997

Molecular and Cellular Biology

Self Cite

116

View full text Add to dashboard Cite

show abstract

“…The SRP54 gene encodes the Y. lipolytica homolog of the Srp54p component of the signal recognition particle involved in protein translocation across the ER membrane (24), while the PEX1 and PEX6 genes encode members of the AAA family of N-ethylmaleimide-sensitive fusion protein-like ATPases that are essential not only for peroxisome biogenesis but also for homo-and heterotypic fusion events required for the assembly of other organelles (46). While previous morphological studies of mammalian and yeast cells have shown a close association between peroxisomes and the ER (for reviews, see references 15 and 22) and recent genetic and biochemical data have suggested a dual role for the ER in supplying phospholipids for the formation of the peroxisomal membrane (45) and in protein trafficking to peroxisomes (2,4,13,46,50,51), this study provides the first genetic evidence that the ER is required for the assembly of functionally intact peroxisomes in Y. lipolytica. A requirement for the ER in peroxisome assembly is also supported by our data showing that not only Pex1p and Pex6p, but also other peroxins essential for peroxisome biogenesis in Y. lipolytica, are required for the exit of secretory proteins from the ER and for their delivery to the extracellular medium (46; this study).…”

Section: Discussionmentioning

confidence: 99%

“…Recent data have suggested a dual role for the ER in peroxisome biogenesis in supplying phospholipid for the formation of the peroxisomal membrane (45) and in protein trafficking to peroxisomes (2,4,13,50,51). We have applied a combined genetic, biochemical, and morphological approach to study the importance of the ER for peroxisome biogenesis in Y. lipolytica.…”

mentioning

confidence: 99%

Mutants of the Yeast Yarrowia lipolyticaDefective in Protein Exit from the Endoplasmic Reticulum Are Also Defective in Peroxisome Biogenesis

Titorenko

Rachubinski

1998

Molecular and Cellular Biology

158

127

View full text Add to dashboard Cite

Mutations in the SEC238 and SRP54 genes of the yeast Yarrowia lipolytica not only cause temperature-sensitive defects in the exit of the precursor form of alkaline extracellular protease and of other secretory proteins from the endoplasmic reticulum and in protein secretion but also lead to temperature-sensitive growth in oleic acid-containing medium, the metabolism of which requires the assembly of functionally intact peroxisomes. The sec238A and srp54KO mutations at the restrictive temperature significantly reduce the size and number of peroxisomes, affect the import of peroxisomal matrix and membrane proteins into the organelle, and significantly delay, but do not prevent, the exit of two peroxisomal membrane proteins, Pex2p and Pex16p, from the endoplasmic reticulum en route to the peroxisomal membrane. Mutations in the PEX1 and PEX6 genes, which encode members of the AAA family of N-ethylmaleimide-sensitive fusion protein-like ATPases, not only affect the exit of precursor forms of secretory proteins from the endoplasmic reticulum but also prevent the exit of the peroxisomal membrane proteins Pex2p and Pex16p from the endoplasmic reticulum and cause the accumulation of an extensive network of endoplasmic reticulum membranes. None of the peroxisomal matrix proteins tested associated with the endoplasmic reticulum in sec238A, srp54KO, pex1-1, and pex6KO mutant cells. Our data provide evidence that the endoplasmic reticulum is required for peroxisome biogenesis and suggest that in Y. lipolytica, the trafficking of some membrane proteins, but not matrix proteins, to the peroxisome occurs via the endoplasmic reticulum, results in their glycosylation within the lumen of the endoplasmic reticulum, does not involve transport through the Golgi, and requires the products encoded by the SEC238, SRP54, PEX1, and PEX6 genes.One of the hallmarks of eukaryotic cells is the coexistence of functionally distinct subcellular organelles (compartments), with each organelle possessing a specific set of enzymes required for its particular metabolic role. One organelle, the peroxisome, is present in most eukaryotic cells (22). Peroxisomes compartmentalize more than 50 enzymes involved in different metabolic functions, including the ␤-oxidation of fatty acids and the decomposition of H 2 O 2 by catalase (42, 49). The importance of peroxisomes for normal human development and physiology is demonstrated by the lethality of various peroxisome biogenesis disorders (23).Changes in the abundance and composition of peroxisomes in response to changes in environmental conditions must be coordinated with the biogenesis and functioning of other organelles in order to achieve an overall balance in cellular function. An example of such interorganellar communication is the tripartite path of communication among mitochondria, the nucleus, and peroxisomes, which regulates the expression of genes encoding peroxisomal proteins (6, 32). Some peroxisomal proteins may also play an important role in the biogenesis of other organelles. The peroxisome-ass...

show abstract

“…Microinjection of fluorochromes 1 , antibodies 2-4 , proteins [5][6][7] , and genetic material [8][9][10] is widely practiced by biologists despite disadvantages associated with insertion of microcapillaries. In small cells, sealing of the plasma membrane around the tip (diameter, ∼1 µm) is often incomplete.…”

Section: Researchmentioning

confidence: 99%

A galinstan expansion femtosyringe for microinjection of eukaryotic organelles and prokaryotes

et al. 1999

View full text Add to dashboard Cite

A galinstan expansion femtosyringe enables femtoliter to attoliter samples to be introduced into prokaryotes and subcellular compartments of eukaryotes. The method uses heat-induced expansion of galinstan (a liquid metal alloy of gallium, indium, and tin) within a glass syringe to expel samples through a tip diameter of about 0.1 microm. The narrow tip inflicts less damage than conventional capillaries, and the heat-induced expansion of the galinstan allows fine control over the rate of injection. We demonstrate injection of Lucifer Yellow and Lucifer Yellow-dextran conjugates into cyanobacteria, and into nuclei and chloroplasts of higher organisms. Injection of a plasmid containing the bla gene into the cyanobacterium Phormidium laminosum resulted in transformed ampicillin-resistant cultures. Green fluorescent protein was expressed in attached leaves of tobacco and Vicia faba following injection of DNA containing its gene into individual chloroplasts.

show abstract

Transport of microinjected alcohol oxidase from Pichia pastoris into vesicles in mammalian cells: involvement of the peroxisomal targeting signal.

Abstract: Abstract. This report describes the microinjection of a purified peroxisomal protein, alcohol oxidase, from

Cited by 31 publications

References 41 publications

Four Distinct Secretory Pathways Serve Protein Secretion, Cell Surface Growth, and Peroxisome Biogenesis in the Yeast Yarrowia lipolytica

Four Distinct Secretory Pathways Serve Protein Secretion, Cell Surface Growth, and Peroxisome Biogenesis in the Yeast Yarrowia lipolytica

Mutants of the Yeast Yarrowia lipolyticaDefective in Protein Exit from the Endoplasmic Reticulum Are Also Defective in Peroxisome Biogenesis

A galinstan expansion femtosyringe for microinjection of eukaryotic organelles and prokaryotes

Contact Info

Product

Resources

About