Sorry, you need to enable JavaScript to visit this website.

Nickel and low CO2-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters

TitleNickel and low CO2-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2011
AuthorsFerrante, P., Diener D.R., Rosenbaum J.L., and Giuliano Giovanni
JournalBMC Plant Biology
Volume11
ISSN14712229
Keywordsarticle, Blotting, Carbon dioxide, Chlamydomonas, Chlamydomonas reinhardtii, Chlorophyta, drug effect, Flagella, flagellum, gene expression regulation, Genetic, genetic complementation, Genetic Complementation Test, genetic transformation, genetics, Kinetics, metabolism, Movement, movement (physiology), mutation, Nickel, Phenotype, physiology, Plant, Plant Proteins, promoter region, Promoter Regions, Reverse Transcriptase Polymerase Chain Reaction, reverse transcription polymerase chain reaction, Time, Time Factors, Transformation, transgene, Transgenes, vegetable protein, Western, Western blotting
Abstract

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 CO2-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 CO2conditions, 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. © 2011 Ferrante et al; licensee BioMed Central Ltd.

Notes

cited By 11

URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-78951468807&doi=10.1186%2f1471-2229-11-22&partnerID=40&md5=696980e8b2492dea7d31c3944cf4b652
DOI10.1186/1471-2229-11-22
Citation KeyFerrante2011