title: Δ(1)-pyrroline-5-carboxylate/glutamate biogenesis is required for fungal virulence and sporulation.
abstract: Proline dehydrogenase (Prodh) and Δ(1)-pyrroline-5-carboxylate dehydrogenase (P5Cdh) are two key enzymes in the cellular biogenesis of glutamate. Recombinant Prodh and P5Cdh proteins of the chestnut blight fungus Cryphonectria parasitica were investigated and showed activity in in vitro assays. Additionally, the C. parasitica Prodh and P5Cdh genes were able to complement the Saccharomyces cerevisiae put1 and put2 null mutants, respectively, to allow these proline auxotrophic yeast mutants to grow on media with proline as the sole source of nitrogen. Deletion of the Prodh gene in C. parasitica resulted in hypovirulence and a lower level of sporulation, whereas deletion of P5Cdh resulted in hypovirulence though no effect on sporulation; both Δprodh and Δp5cdh mutants were unable to grow on minimal medium with proline as the sole nitrogen source. In a wild-type strain, the intracellular level of proline and the activity of Prodh and P5Cdh increased after supplementation of exogenous proline, though the intracellular Δ(1)-pyrroline-5-carboxylate (P5C) content remained unchanged. Prodh and P5Cdh were both transcriptionally down-regulated in cells infected with hypovirus. The disruption of other genes with products involved in the conversion of arginine to ornithine, ornithine and glutamate to P5C, and P5C to proline in the cytosol did not appear to affect virulence; however, asexual sporulation was reduced in the Δpro1 and Δpro2 mutants. Taken together, our results showed that Prodh, P5Cdh and related mitochondrial functions are essential for virulence and that proline/glutamate pathway components may represent down-stream targets of hypovirus regulation in C. parasitica.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, College of Life Science and Technology, Guangxi University, Nanning, China.
authors.forename: Ziting
authors.initials: Z
authors.lastname: Yao
authors.forename: Chengwu
authors.initials: C
authors.lastname: Zou
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.forename: Jinzi
authors.initials: J
authors.lastname: Wang
authors.forename: Lidan
authors.initials: L
authors.lastname: Lu
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Baoshan
authors.initials: B
authors.lastname: Chen
mesh_terms: D050842:1-Pyrroline-5-Carboxylate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001203:Ascomycota / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology* / Q000821:virology; D031256:Eleocharis / Q000469:parasitology*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D018698:Glutamic Acid / Q000378:metabolism*; D010802:Phylogeny; D011394:Proline Oxidase / Q000235:genetics / Q000378:metabolism; D011758:Pyrroles / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013172:Spores, Fungal / Q000235:genetics / Q000378:metabolism; D014774:Virulence
chemical_list: D005656:Fungal Proteins; D011758:Pyrroles; D011994:Recombinant Proteins; C015485:delta-1-pyrroline-5-carboxylate; D018698:Glutamic Acid; D050842:1-Pyrroline-5-Carboxylate Dehydrogenase; D011394:Proline Oxidase
references.citation: Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):305-9
references.pmid: 7816838
references.citation: Mol Plant Microbe Interact. 2008 Dec;21(12):1582-90
references.pmid: 18986254
references.citation: J Bacteriol. 2000 Jun;182(11):3158-64
references.pmid: 10809695
references.citation: Microbiology (Reading). 2005 May;151(Pt 5):1349-1358
references.pmid: 15870445
references.citation: Fungal Genet Biol. 2013 Feb;51:60-71
references.pmid: 23084963
references.citation: Genetics. 1982 Sep;102(1):25-8
references.pmid: 17246094
references.citation: Science. 1992 Aug 7;257(5071):800-3
references.pmid: 1496400
references.citation: Mol Microbiol. 2004 Mar;51(5):1267-77
references.pmid: 14982623
references.citation: Biochem Genet. 2007 Jun;45(5-6):487-506
references.pmid: 17497221
references.citation: Fungal Genet Biol. 2006 Sep;43(9):605-17
references.pmid: 16731015
references.citation: J Bacteriol. 1987 Nov;169(11):5324-6
references.pmid: 3667533
references.citation: Microbiology (Reading). 2009 Dec;155(Pt 12):3913-3921
references.pmid: 19589830
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12616-21
references.pmid: 15308773
references.citation: J Proteomics. 2013 Jan 14;78:221-30
references.pmid: 22954595
references.citation: Mol Plant Pathol. 2012 Apr;13(3):240-50
references.pmid: 21895942
references.citation: Mol Plant Microbe Interact. 1997 Nov;10(8):984-93
references.pmid: 9353946
references.citation: J Bacteriol. 1992 Aug;174(15):5176
references.pmid: 1352771
references.citation: Mol Cell Biol. 1986 Jul;6(7):2638-45
references.pmid: 3537723
references.citation: Eukaryot Cell. 2004 Oct;3(5):1227-32
references.pmid: 15470251
references.citation: Curr Genet. 2003 Apr;43(1):24-33
references.pmid: 12684842
references.citation: J Bacteriol. 1979 Nov;140(2):498-503
references.pmid: 387737
references.citation: Mol Plant Pathol. 2011 Apr;12(3):239-46
references.pmid: 21355996
references.citation: Carcinogenesis. 2010 Mar;31(3):446-54
references.pmid: 19942609
references.citation: EMBO J. 1992 Feb;11(2):473-7
references.pmid: 1537330
references.citation: Cancer Res. 2009 Aug 15;69(16):6414-22
references.pmid: 19654292
references.citation: FEMS Microbiol Lett. 2000 Mar 1;184(1):103-8
references.pmid: 10689174
references.citation: FEMS Microbiol Lett. 2012 Nov;336(1):64-72
references.pmid: 22889301
references.citation: Eukaryot Cell. 2004 Dec;3(6):1454-63
references.pmid: 15590820
references.citation: Mol Gen Genet. 1996 Dec 13;253(3):334-41
references.pmid: 9003320
references.citation: FEBS Lett. 1999 Nov 19;461(3):205-10
references.pmid: 10567698
references.citation: Yeast. 1995 Apr 15;11(4):355-60
references.pmid: 7785336
references.citation: Plant Cell. 1996 Aug;8(8):1323-35
references.pmid: 8776899
references.citation: Fungal Genet Biol. 2006 May;43(5):326-36
references.pmid: 16540355
references.citation: Mol Cell Biol. 1987 Dec;7(12):4431-40
references.pmid: 3125423
references.citation: J Plant Physiol. 2011 Sep 1;168(13):1588-97
references.pmid: 21481968
references.citation: Eur J Biochem. 1987 May 4;164(3):601-6
references.pmid: 3552672
references.citation: Curr Genet. 2008 Jan;53(1):59-66
references.pmid: 17972079
references.citation: J Biosci Bioeng. 2005 Nov;100(5):538-44
references.pmid: 16384793
references.citation: J Bacteriol. 1987 Dec;169(12):5364-72
references.pmid: 2824433
references.citation: Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5935-9
references.pmid: 11607549
references.citation: Plant J. 2001 Aug;27(4):345-56
references.pmid: 11532180
references.citation: Appl Microbiol Biotechnol. 2008 May;79(2):273-83
references.pmid: 18351334
references.citation: J Mol Biol. 2012 Jul 13;420(3):176-89
references.pmid: 22516612
references.citation: J Virol. 1993 Nov;67(11):6513-21
references.pmid: 8411354
references.citation: Nat Rev Microbiol. 2005 Aug;3(8):632-42
references.pmid: 16064055
references.citation: BMC Plant Biol. 2010 Apr 19;10:70
references.pmid: 20403182
references.citation: Phytopathology. 2010 Oct;100(10):1100-10
references.pmid: 20839945
references.citation: Genetics. 2012 Jan;190(1):113-27
references.pmid: 22021387
references.citation: Genetics. 1996 Apr;142(4):1069-82
references.pmid: 8846888
references.citation: Plant Cell. 2004 Dec;16(12):3413-25
references.pmid: 15548746
references.citation: Curr Microbiol. 2008 Apr;56(4):363-70
references.pmid: 18172716
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14122-7
references.pmid: 11038529
references.citation: J Biol Chem. 2010 Aug 20;285(34):26685-96
references.pmid: 20558740
references.citation: Can J Microbiol. 2012 Jul;58(7):863-71
references.pmid: 22716047
references.citation: Appl Environ Microbiol. 2003 Mar;69(3):1623-8
references.pmid: 12620851
references.citation: Plant Cell Physiol. 2003 May;44(5):541-8
references.pmid: 12773641
references.citation: Appl Environ Microbiol. 1998 Aug;64(8):2988-94
references.pmid: 9687462
references.citation: J Biol Chem. 2007 Mar 30;282(13):9581-9590
references.pmid: 17244616
references.citation: Fungal Genet Biol. 2004 Nov;41(11):973-81
references.pmid: 15465386
references.citation: J Med Entomol. 2005 May;42(3):346-51
references.pmid: 15962786
references.citation: Appl Environ Microbiol. 2006 Jun;72(6):4001-6
references.pmid: 16751508
references.citation: Appl Microbiol Biotechnol. 2012 Jan;93(1):83-93
references.pmid: 22089387
references.citation: Fungal Biol. 2012 Mar;116(3):363-78
references.pmid: 22385619
references.citation: J Biol Chem. 1978 Sep 10;253(17):5997-6001
references.pmid: 355248
references.citation: J Biol Chem. 2009 Sep 25;284(39):26482-92
references.pmid: 19635803
references.citation: J Mol Biol. 1994 Nov 11;243(5):950-6
references.pmid: 7966312