Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae.
Both the yeast nascent polypeptide-associated complex (NAC) and the Hsp40/70-based chaperone system RAC-Ssb are systems tethered to the ribosome to assist cotranslational processes such as folding of nascent polypeptides. While loss of NAC does not cause phenotypic changes in yeast, the simultaneous...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4664479?pdf=render |
| _version_ | 1818214604232196096 |
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| author | Ann-Kathrin Ott Lisa Locher Miriam Koch Elke Deuerling |
| author_facet | Ann-Kathrin Ott Lisa Locher Miriam Koch Elke Deuerling |
| author_sort | Ann-Kathrin Ott |
| collection | DOAJ |
| description | Both the yeast nascent polypeptide-associated complex (NAC) and the Hsp40/70-based chaperone system RAC-Ssb are systems tethered to the ribosome to assist cotranslational processes such as folding of nascent polypeptides. While loss of NAC does not cause phenotypic changes in yeast, the simultaneous deletion of genes coding for NAC and the chaperone Ssb (nacΔssbΔ) leads to strongly aggravated defects compared to cells lacking only Ssb, including impaired growth on plates containing L-canavanine or hygromycin B, aggregation of newly synthesized proteins and a reduced translational activity due to ribosome biogenesis defects. In this study, we dissected the functional properties of the individual NAC-subunits (α-NAC, β-NAC and β'-NAC) and of different NAC heterodimers found in yeast (αβ-NAC and αβ'-NAC) by analyzing their capability to complement the pleiotropic phenotype of nacΔssbΔ cells. We show that the abundant heterodimer αβ-NAC but not its paralogue αβ'-NAC is able to suppress all phenotypic defects of nacΔssbΔ cells including global protein aggregation as well as translation and growth deficiencies. This suggests that αβ-NAC and αβ'-NAC are functionally distinct from each other. The function of αβ-NAC strictly depends on its ribosome association and on its high level of expression. Expression of individual β-NAC, β'-NAC or α-NAC subunits as well as αβ'-NAC ameliorated protein aggregation in nacΔssbΔ cells to different extents while only β-NAC was able to restore growth defects suggesting chaperoning activities for β-NAC sufficient to decrease the sensitivity of nacΔssbΔ cells against L-canavanine or hygromycin B. Interestingly, deletion of the ubiquitin-associated (UBA)-domain of the α-NAC subunit strongly enhanced the aggregation preventing activity of αβ-NAC pointing to a negative regulatory role of this domain for the NAC chaperone activity in vivo. |
| first_indexed | 2024-12-12T06:22:49Z |
| format | Article |
| id | doaj.art-30be15e9476643bca56f16fdde50c6b8 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T06:22:49Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-30be15e9476643bca56f16fdde50c6b82022-12-22T00:34:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011011e014345710.1371/journal.pone.0143457Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae.Ann-Kathrin OttLisa LocherMiriam KochElke DeuerlingBoth the yeast nascent polypeptide-associated complex (NAC) and the Hsp40/70-based chaperone system RAC-Ssb are systems tethered to the ribosome to assist cotranslational processes such as folding of nascent polypeptides. While loss of NAC does not cause phenotypic changes in yeast, the simultaneous deletion of genes coding for NAC and the chaperone Ssb (nacΔssbΔ) leads to strongly aggravated defects compared to cells lacking only Ssb, including impaired growth on plates containing L-canavanine or hygromycin B, aggregation of newly synthesized proteins and a reduced translational activity due to ribosome biogenesis defects. In this study, we dissected the functional properties of the individual NAC-subunits (α-NAC, β-NAC and β'-NAC) and of different NAC heterodimers found in yeast (αβ-NAC and αβ'-NAC) by analyzing their capability to complement the pleiotropic phenotype of nacΔssbΔ cells. We show that the abundant heterodimer αβ-NAC but not its paralogue αβ'-NAC is able to suppress all phenotypic defects of nacΔssbΔ cells including global protein aggregation as well as translation and growth deficiencies. This suggests that αβ-NAC and αβ'-NAC are functionally distinct from each other. The function of αβ-NAC strictly depends on its ribosome association and on its high level of expression. Expression of individual β-NAC, β'-NAC or α-NAC subunits as well as αβ'-NAC ameliorated protein aggregation in nacΔssbΔ cells to different extents while only β-NAC was able to restore growth defects suggesting chaperoning activities for β-NAC sufficient to decrease the sensitivity of nacΔssbΔ cells against L-canavanine or hygromycin B. Interestingly, deletion of the ubiquitin-associated (UBA)-domain of the α-NAC subunit strongly enhanced the aggregation preventing activity of αβ-NAC pointing to a negative regulatory role of this domain for the NAC chaperone activity in vivo.http://europepmc.org/articles/PMC4664479?pdf=render |
| spellingShingle | Ann-Kathrin Ott Lisa Locher Miriam Koch Elke Deuerling Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. PLoS ONE |
| title | Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. |
| title_full | Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. |
| title_fullStr | Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. |
| title_full_unstemmed | Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. |
| title_short | Functional Dissection of the Nascent Polypeptide-Associated Complex in Saccharomyces cerevisiae. |
| title_sort | functional dissection of the nascent polypeptide associated complex in saccharomyces cerevisiae |
| url | http://europepmc.org/articles/PMC4664479?pdf=render |
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