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Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters

BACKGROUND: Chlamydomonas reinhardtii is a model system for the biology of unicellular green algae. Chemically regulated promoters, such as the nickel-inducible CYC6 or the low CO(2)-inducible CAH1 promoter, may prove useful for expressing, at precise times during its cell cycle, proteins with relev...

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Autores principales: Ferrante, Paola, Diener, Dennis R, Rosenbaum, Joel L, Giuliano, Giovanni
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3038898/
https://www.ncbi.nlm.nih.gov/pubmed/21266063
http://dx.doi.org/10.1186/1471-2229-11-22
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author Ferrante, Paola
Diener, Dennis R
Rosenbaum, Joel L
Giuliano, Giovanni
author_facet Ferrante, Paola
Diener, Dennis R
Rosenbaum, Joel L
Giuliano, Giovanni
author_sort Ferrante, Paola
collection PubMed
description BACKGROUND: Chlamydomonas reinhardtii is a model system for the biology of unicellular green algae. Chemically regulated promoters, such as the nickel-inducible CYC6 or the low CO(2)-inducible CAH1 promoter, may prove useful for expressing, at precise times during its cell cycle, proteins with relevant biological functions, or complementing mutants in genes encoding such proteins. To this date, this has not been reported for the above promoters. RESULTS: We fused the CYC6 and CAH1 promoters to an HA-tagged RSP3 gene, encoding a protein of the flagellar radial spoke complex. The constructs were used for chemically regulated complementation of the pf14 mutant, carrying an ochre mutation in the RSP3 gene. 7 to 8% of the transformants showed cells with restored motility after induction with nickel or transfer to low CO(2 )conditions, but not in non-inducing conditions. Maximum complementation (5% motile cells) was reached with very different kinetics (5-6 hours for CAH1, 48 hours for CYC6). The two inducible promoters drive much lower levels of RSP3 protein expression than the constitutive PSAD promoter, which shows almost complete rescue of motility. CONCLUSIONS: To our knowledge, this is the first example of the use of the CYC6 or CAH1 promoters to perform a chemically regulated complementation of a Chlamydomonas mutant. Based on our data, the CYC6 and CAH1 promoters should be capable of fully complementing mutants in genes whose products exert their biological activity at low concentrations.
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spelling pubmed-30388982011-02-15 Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters Ferrante, Paola Diener, Dennis R Rosenbaum, Joel L Giuliano, Giovanni BMC Plant Biol Research Article BACKGROUND: Chlamydomonas reinhardtii is a model system for the biology of unicellular green algae. Chemically regulated promoters, such as the nickel-inducible CYC6 or the low CO(2)-inducible CAH1 promoter, may prove useful for expressing, at precise times during its cell cycle, proteins with relevant biological functions, or complementing mutants in genes encoding such proteins. To this date, this has not been reported for the above promoters. RESULTS: We fused the CYC6 and CAH1 promoters to an HA-tagged RSP3 gene, encoding a protein of the flagellar radial spoke complex. The constructs were used for chemically regulated complementation of the pf14 mutant, carrying an ochre mutation in the RSP3 gene. 7 to 8% of the transformants showed cells with restored motility after induction with nickel or transfer to low CO(2 )conditions, but not in non-inducing conditions. Maximum complementation (5% motile cells) was reached with very different kinetics (5-6 hours for CAH1, 48 hours for CYC6). The two inducible promoters drive much lower levels of RSP3 protein expression than the constitutive PSAD promoter, which shows almost complete rescue of motility. CONCLUSIONS: To our knowledge, this is the first example of the use of the CYC6 or CAH1 promoters to perform a chemically regulated complementation of a Chlamydomonas mutant. Based on our data, the CYC6 and CAH1 promoters should be capable of fully complementing mutants in genes whose products exert their biological activity at low concentrations. BioMed Central 2011-01-25 /pmc/articles/PMC3038898/ /pubmed/21266063 http://dx.doi.org/10.1186/1471-2229-11-22 Text en Copyright ©2011 Ferrante et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ferrante, Paola
Diener, Dennis R
Rosenbaum, Joel L
Giuliano, Giovanni
Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title_full Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title_fullStr Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title_full_unstemmed Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title_short Nickel and low CO(2)-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
title_sort nickel and low co(2)-controlled motility in chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3038898/
https://www.ncbi.nlm.nih.gov/pubmed/21266063
http://dx.doi.org/10.1186/1471-2229-11-22
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