RTA1 Is Involved in Resistance to 7-Aminocholesterol and Secretion of Fungal Proteins in Cryptococcus neoformans
Cryptococcus neoformans (Cn) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence factors, such as the enzyme laccase and its product melanin, phospholipase, and capsular polysaccharide have been identified. During a screen of knock...
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| Published in: | Pathogens (Basel) Vol. 11; no. 11; p. 1239 |
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| Main Authors: | , , , , , , , , |
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| Abstract | Cryptococcus neoformans (Cn) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence factors, such as the enzyme laccase and its product melanin, phospholipase, and capsular polysaccharide have been identified. During a screen of knockout mutants, the gene resistance to aminocholesterol 1 (RTA1) was identified, the function of which is currently unknown in Cn. Rta1 homologs in S. cerevisiae belong to a lipid-translocating exporter family of fungal proteins with transmembrane regions and confer resistance to the antimicrobial agent 7-aminocholesterol when overexpressed. To determine the role of RTA1 in Cn, the knock-out (rta1Δ) and reconstituted (rta1Δ+RTA1) strains were created and phenotypically tested. RTA1 was involved in resistance to 7-aminocholesterol, and also in exocyst complex component 3 (Sec6)-mediated secretion of urease, laccase, and the major capsule component, glucuronoxylomannan (GXM), which coincided with significantly smaller capsules in the rta1Δ and rta1Δ+RTA1 strains compared to the wild-type H99 strain. Furthermore, RTA1 expression was reduced in a secretory 14 mutant (sec14Δ) and increased in an RNAi Sec6 mutant. Transmission electron microscopy demonstrated vesicle accumulation inside the rta1Δ strain, predominantly near the cell membrane. Given that Rta1 is likely to be a transmembrane protein located at the plasma membrane, these data suggest that Rta1 may be involved in both secretion of various fungal virulence factors and resistance to 7-aminocholesterol in Cn. |
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| AbstractList | Cryptococcus neoformans
(
Cn
) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various
Cn
virulence factors, such as the enzyme laccase and its product melanin, phospholipase, and capsular polysaccharide have been identified. During a screen of knockout mutants, the gene resistance to aminocholesterol 1 (
RTA1
) was identified, the function of which is currently unknown in
Cn
. Rta1 homologs in
S. cerevisiae
belong to a lipid-translocating exporter family of fungal proteins with transmembrane regions and confer resistance to the antimicrobial agent 7-aminocholesterol when overexpressed. To determine the role of
RTA1
in
Cn
, the knock-out (
rta1Δ
) and reconstituted (
rta1Δ+RTA1
) strains were created and phenotypically tested.
RTA1
was involved in resistance to 7-aminocholesterol, and also in exocyst complex component 3 (Sec6)-mediated secretion of urease, laccase, and the major capsule component, glucuronoxylomannan (GXM), which coincided with significantly smaller capsules in the
rta1Δ
and
rta1Δ+RTA1
strains compared to the wild-type H99 strain. Furthermore,
RTA1
expression was reduced in a secretory 14 mutant (
sec14Δ
) and increased in an RNAi Sec6 mutant. Transmission electron microscopy demonstrated vesicle accumulation inside the
rta1Δ
strain, predominantly near the cell membrane. Given that Rta1 is likely to be a transmembrane protein located at the plasma membrane, these data suggest that Rta1 may be involved in both secretion of various fungal virulence factors and resistance to 7-aminocholesterol in
Cn
. Cryptococcus neoformans (Cn) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence factors, such as the enzyme laccase and its product melanin, phospholipase, and capsular polysaccharide have been identified. During a screen of knockout mutants, the gene resistance to aminocholesterol 1 (RTA1) was identified, the function of which is currently unknown in Cn. Rta1 homologs in S. cerevisiae belong to a lipid-translocating exporter family of fungal proteins with transmembrane regions and confer resistance to the antimicrobial agent 7-aminocholesterol when overexpressed. To determine the role of RTA1 in Cn, the knock-out (rta1Δ) and reconstituted (rta1Δ+RTA1) strains were created and phenotypically tested. RTA1 was involved in resistance to 7-aminocholesterol, and also in exocyst complex component 3 (Sec6)-mediated secretion of urease, laccase, and the major capsule component, glucuronoxylomannan (GXM), which coincided with significantly smaller capsules in the rta1Δ and rta1Δ+RTA1 strains compared to the wild-type H99 strain. Furthermore, RTA1 expression was reduced in a secretory 14 mutant (sec14Δ) and increased in an RNAi Sec6 mutant. Transmission electron microscopy demonstrated vesicle accumulation inside the rta1Δ strain, predominantly near the cell membrane. Given that Rta1 is likely to be a transmembrane protein located at the plasma membrane, these data suggest that Rta1 may be involved in both secretion of various fungal virulence factors and resistance to 7-aminocholesterol in Cn. |
| Author | Smith-Peavler, Emily S. Bowring, Bethany G. Prabu, Moses M. Onumajuru, Adejumoke Mary Djordjevic, Julianne T. Patel, Ronakkumar Brunel, Jean Michel McClelland, Erin E. Miller, Joyce L. |
| AuthorAffiliation | 1 Department of Biology, Middle Tennessee State University, Murfreesboro, TN 37132, USA 5 UMR_MD1 “Membranes et Cibles Thérapeutiques”, Faculté de Pharmacie, Aix Marseille Universite, INSERM, SSA, MCT, 13385 Marseille, France 8 Biomedical Sciences Division, Marian University College of Osteopathic Medicine, Indianapolis, IN 46234, USA 6 Sydney Institute for Infectious Diseases, University of Sydney, Sydney, NSW 2145, Australia 7 M&P Associates, Inc., Murfreesboro, TN 37129, USA 4 MTSU Interdisciplinary Microanalysis and Imaging Center, Middle Tennessee State University, Murfreesboro, TN 37132, USA 3 Centre for Infectious Diseases and Microbiology, The Westmead Institute for Medical Research, Sydney, NSW 2145, Australia 2 Otsuka Pharmaceutical, 508 Carnegie Center Dr., Princeton, NJ 08540, USA |
| AuthorAffiliation_xml | – name: 6 Sydney Institute for Infectious Diseases, University of Sydney, Sydney, NSW 2145, Australia – name: 2 Otsuka Pharmaceutical, 508 Carnegie Center Dr., Princeton, NJ 08540, USA – name: 3 Centre for Infectious Diseases and Microbiology, The Westmead Institute for Medical Research, Sydney, NSW 2145, Australia – name: 8 Biomedical Sciences Division, Marian University College of Osteopathic Medicine, Indianapolis, IN 46234, USA – name: 5 UMR_MD1 “Membranes et Cibles Thérapeutiques”, Faculté de Pharmacie, Aix Marseille Universite, INSERM, SSA, MCT, 13385 Marseille, France – name: 7 M&P Associates, Inc., Murfreesboro, TN 37129, USA – name: 4 MTSU Interdisciplinary Microanalysis and Imaging Center, Middle Tennessee State University, Murfreesboro, TN 37132, USA – name: 1 Department of Biology, Middle Tennessee State University, Murfreesboro, TN 37132, USA |
| Author_xml | – sequence: 1 givenname: Emily S. surname: Smith-Peavler fullname: Smith-Peavler, Emily S. – sequence: 2 givenname: Ronakkumar surname: Patel fullname: Patel, Ronakkumar – sequence: 3 givenname: Adejumoke Mary orcidid: 0000-0002-8963-7192 surname: Onumajuru fullname: Onumajuru, Adejumoke Mary – sequence: 4 givenname: Bethany G. surname: Bowring fullname: Bowring, Bethany G. – sequence: 5 givenname: Joyce L. surname: Miller fullname: Miller, Joyce L. – sequence: 6 givenname: Jean Michel orcidid: 0000-0002-9355-8980 surname: Brunel fullname: Brunel, Jean Michel – sequence: 7 givenname: Julianne T. orcidid: 0000-0003-4207-4115 surname: Djordjevic fullname: Djordjevic, Julianne T. – sequence: 8 givenname: Moses M. surname: Prabu fullname: Prabu, Moses M. – sequence: 9 givenname: Erin E. orcidid: 0000-0001-7781-3542 surname: McClelland fullname: McClelland, Erin E. |
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| Keywords | secretion extracellular vesicles Cryptococcus neoformans RTA1 |
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| Snippet | Cryptococcus neoformans (Cn) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence factors,... Cryptococcus neoformans ( Cn ) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence... |
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| Title | RTA1 Is Involved in Resistance to 7-Aminocholesterol and Secretion of Fungal Proteins in Cryptococcus neoformans |
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