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Function of a p24 Heterodimer in Morphogenesis and Protein Transport in Penicillium oxalicum.

Wang F, Liu K, Han L, Jiang B, Wang M, Fang X - Sci Rep (2015)

Bottom Line: The results suggested that the p24 heterodimer mediates protein transport, particularly that of cellobiohydrolase.These results suggest that the p24 heterodimer participates in morphogenesis and protein transport.Compared with P. oxalicum Δerp, a greater number of cellular physiological pathways were impaired in ΔerpΔpδ.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Jinan, Shandong, China.

ABSTRACT
The lignocellulose degradation capacity of filamentous fungi has been widely studied because of their cellulase hypersecretion. The p24 proteins in eukaryotes serve important functions in this secretory pathway. However, little is known about the functions of the p24 proteins in filamentous fungi. In this study, four p24 proteins were identified in Penicillium oxalicum. Six p24 double-deletion strains were constructed, and further studies were carried out with the ΔerpΔpδ strain. The experimental results suggested that Erp and Pδ form a p24 heterodimer in vivo. This p24 heterodimer participates in important morphogenetic events, including sporulation, hyphal growth, and lateral branching. The results suggested that the p24 heterodimer mediates protein transport, particularly that of cellobiohydrolase. Analysis of the intracellular proteome revealed that the ΔerpΔpδ double mutant is under secretion stress due to attempts to remove proteins that are jammed in the endomembrane system. These results suggest that the p24 heterodimer participates in morphogenesis and protein transport. Compared with P. oxalicum Δerp, a greater number of cellular physiological pathways were impaired in ΔerpΔpδ. This finding may provide new insights into the secretory pathways of filamentous fungi.

No MeSH data available.


Related in: MedlinePlus

Erp and Pδ Homologs in P. oxalicum 114-2.Panel a, phylogenetic tree of certain p24 proteins from eukaryotic species. Black rectangles represent PDE_09471, PDE_08336, PDE_00613 and PDE_03657. H, Homo sapiens; D, Drosophila melanogaster; S, Saccharomyces cerevisiae; X, Xenopus tropicalis; P, Pan troglodytes; A, Arabidopsis thaliana. Protein accession numbers are listed in Supplementary Information S1 online. Panel b, structural diagram of the p24 heterodimer, which is composed of Erp and Pδ. White represents Pδ, and black represents Erp. Each protein domain is marked with a bracket. The di-aromatic/large hydrophobic and di-basic motifs are highlighted with rectangles.
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f1: Erp and Pδ Homologs in P. oxalicum 114-2.Panel a, phylogenetic tree of certain p24 proteins from eukaryotic species. Black rectangles represent PDE_09471, PDE_08336, PDE_00613 and PDE_03657. H, Homo sapiens; D, Drosophila melanogaster; S, Saccharomyces cerevisiae; X, Xenopus tropicalis; P, Pan troglodytes; A, Arabidopsis thaliana. Protein accession numbers are listed in Supplementary Information S1 online. Panel b, structural diagram of the p24 heterodimer, which is composed of Erp and Pδ. White represents Pδ, and black represents Erp. Each protein domain is marked with a bracket. The di-aromatic/large hydrophobic and di-basic motifs are highlighted with rectangles.

Mentions: A BlastP search was performed against the P. oxalicum 114-2 genome with Saccharomyces cerevisiae Emp24, Erv25, Erp3, and Erp5 as queries, from which PDE_00613 (5e-46 E-value, 46% identity), PDE_08336 (3e-55 E-value, 50% identity), PDE_09471 (4e-26 E-value, 31% identity), and PDE_03657 (2e-41 E-value, 38% identity) were identified as the best hits, respectively. A BlastP search was also performed against the S. cerevisiae genome with PDE_00613, PDE_08336, PDE_09471, and PDE_03657 as queries, during which the best hits obtained were Emp24 (3e-46 E-value, 46% identity), Erv25 (5e-55 E-value, 50% identity), Erp3 (7e-26 E-value, 50% identity), and Erp5 (3e-41 E-value, 38% identity), respectively. The sequence alignment of these proteins is shown online in Supplementary Information Fig. S1. Phylogenetic analysis of the four proteins in comparison with other p24 homologs suggested that PDE_03657, PDE_00613, PDE_08336, and PDE_09471 belong to the α, β, δ and γ subfamilies, respectively (Fig. 1a). Six p24 protein double deletion strains were constructed and confirmed by PCR. Their intracellular proteins were analysed using 15% SDS-PAGE. The protein bands of the various p24 deletion strains appeared to be different. The strain that lacked PDE_09471 and PDE_08336 exhibited the most significant changes in comparison to the parent strain (see Supplementary Information Fig. S2 online). As a result, PDE_09471 and PDE_08336 were selected for further investigation. PDE_09471 has been described in detail27 and has been termed Erp. The PDE_08336 was termed Pδ to be consistent with the nomenclature of the p24 family. The ORF of pδ is 892 bp long and is interrupted by two introns6. As it is known that members of the p24 family usually share similar topologies29, the structure diagrams of Erp and Pδ (Fig. 1b) were constructed by comparing their sequences with those of other p24 members. The conservative motifs predicted to be involved in COPI and COPII binding are marked with rectangles in Fig. 1b172630.


Function of a p24 Heterodimer in Morphogenesis and Protein Transport in Penicillium oxalicum.

Wang F, Liu K, Han L, Jiang B, Wang M, Fang X - Sci Rep (2015)

Erp and Pδ Homologs in P. oxalicum 114-2.Panel a, phylogenetic tree of certain p24 proteins from eukaryotic species. Black rectangles represent PDE_09471, PDE_08336, PDE_00613 and PDE_03657. H, Homo sapiens; D, Drosophila melanogaster; S, Saccharomyces cerevisiae; X, Xenopus tropicalis; P, Pan troglodytes; A, Arabidopsis thaliana. Protein accession numbers are listed in Supplementary Information S1 online. Panel b, structural diagram of the p24 heterodimer, which is composed of Erp and Pδ. White represents Pδ, and black represents Erp. Each protein domain is marked with a bracket. The di-aromatic/large hydrophobic and di-basic motifs are highlighted with rectangles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493713&req=5

f1: Erp and Pδ Homologs in P. oxalicum 114-2.Panel a, phylogenetic tree of certain p24 proteins from eukaryotic species. Black rectangles represent PDE_09471, PDE_08336, PDE_00613 and PDE_03657. H, Homo sapiens; D, Drosophila melanogaster; S, Saccharomyces cerevisiae; X, Xenopus tropicalis; P, Pan troglodytes; A, Arabidopsis thaliana. Protein accession numbers are listed in Supplementary Information S1 online. Panel b, structural diagram of the p24 heterodimer, which is composed of Erp and Pδ. White represents Pδ, and black represents Erp. Each protein domain is marked with a bracket. The di-aromatic/large hydrophobic and di-basic motifs are highlighted with rectangles.
Mentions: A BlastP search was performed against the P. oxalicum 114-2 genome with Saccharomyces cerevisiae Emp24, Erv25, Erp3, and Erp5 as queries, from which PDE_00613 (5e-46 E-value, 46% identity), PDE_08336 (3e-55 E-value, 50% identity), PDE_09471 (4e-26 E-value, 31% identity), and PDE_03657 (2e-41 E-value, 38% identity) were identified as the best hits, respectively. A BlastP search was also performed against the S. cerevisiae genome with PDE_00613, PDE_08336, PDE_09471, and PDE_03657 as queries, during which the best hits obtained were Emp24 (3e-46 E-value, 46% identity), Erv25 (5e-55 E-value, 50% identity), Erp3 (7e-26 E-value, 50% identity), and Erp5 (3e-41 E-value, 38% identity), respectively. The sequence alignment of these proteins is shown online in Supplementary Information Fig. S1. Phylogenetic analysis of the four proteins in comparison with other p24 homologs suggested that PDE_03657, PDE_00613, PDE_08336, and PDE_09471 belong to the α, β, δ and γ subfamilies, respectively (Fig. 1a). Six p24 protein double deletion strains were constructed and confirmed by PCR. Their intracellular proteins were analysed using 15% SDS-PAGE. The protein bands of the various p24 deletion strains appeared to be different. The strain that lacked PDE_09471 and PDE_08336 exhibited the most significant changes in comparison to the parent strain (see Supplementary Information Fig. S2 online). As a result, PDE_09471 and PDE_08336 were selected for further investigation. PDE_09471 has been described in detail27 and has been termed Erp. The PDE_08336 was termed Pδ to be consistent with the nomenclature of the p24 family. The ORF of pδ is 892 bp long and is interrupted by two introns6. As it is known that members of the p24 family usually share similar topologies29, the structure diagrams of Erp and Pδ (Fig. 1b) were constructed by comparing their sequences with those of other p24 members. The conservative motifs predicted to be involved in COPI and COPII binding are marked with rectangles in Fig. 1b172630.

Bottom Line: The results suggested that the p24 heterodimer mediates protein transport, particularly that of cellobiohydrolase.These results suggest that the p24 heterodimer participates in morphogenesis and protein transport.Compared with P. oxalicum Δerp, a greater number of cellular physiological pathways were impaired in ΔerpΔpδ.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Jinan, Shandong, China.

ABSTRACT
The lignocellulose degradation capacity of filamentous fungi has been widely studied because of their cellulase hypersecretion. The p24 proteins in eukaryotes serve important functions in this secretory pathway. However, little is known about the functions of the p24 proteins in filamentous fungi. In this study, four p24 proteins were identified in Penicillium oxalicum. Six p24 double-deletion strains were constructed, and further studies were carried out with the ΔerpΔpδ strain. The experimental results suggested that Erp and Pδ form a p24 heterodimer in vivo. This p24 heterodimer participates in important morphogenetic events, including sporulation, hyphal growth, and lateral branching. The results suggested that the p24 heterodimer mediates protein transport, particularly that of cellobiohydrolase. Analysis of the intracellular proteome revealed that the ΔerpΔpδ double mutant is under secretion stress due to attempts to remove proteins that are jammed in the endomembrane system. These results suggest that the p24 heterodimer participates in morphogenesis and protein transport. Compared with P. oxalicum Δerp, a greater number of cellular physiological pathways were impaired in ΔerpΔpδ. This finding may provide new insights into the secretory pathways of filamentous fungi.

No MeSH data available.


Related in: MedlinePlus