Flagellar Elongation and Gene Expression in<i>Chlamydomonas reinhardtii</i>

Periz, Goran; Dharia, Darshita; Miller, Steven H.; Keller, Laura R.

doi:10.1128/ec.00167-07

Cited by 16 publications

(15 citation statements)

References 54 publications

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“…Similar treatment of RNAi strains did not show apparent length difference. It has been reported that longer treatment of cells induces formation of bulbed flagella at the flagellar tip, and the flagella begin to curl at the base forming big “bulbs” which are eventually lost forming aflagellate cells [52], [53]. We have confirmed this observation, as shown in Figure 6B and C.…”

Section: Resultssupporting

confidence: 88%

“…Inhibition of GSK3β [52] or adenylate cyclase [51] has been proposed to underlie the effect of LiCl. Treatment of Chlamydomonas wild type cells with 25 mM LiCl induced about 40% increase of flagellar length from 12 µm to 17 µm similarly as reported (Figure 6A) [53]. Similar treatment of RNAi strains did not show apparent length difference.…”

Section: Resultssupporting

confidence: 81%

“…LiCl has been shown to stimulate cilia elongation in vertebrate cells [51] and Chlamydomonas [52], [53] [54],. Inhibition of GSK3β [52] or adenylate cyclase [51] has been proposed to underlie the effect of LiCl.…”

Section: Resultsmentioning

confidence: 99%

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Regulation of Flagellar Biogenesis by a Calcium Dependent Protein Kinase in Chlamydomonas reinhardtii

Liang

Pan

2013

PLoS ONE

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Chlamydomonas reinhardtii, a bi-flagellated green alga, is a model organism for studies of flagella or cilia related activities including cilia-based signaling, flagellar motility and flagellar biogenesis. Calcium has been shown to be a key regulator of these cellular processes whereas the signaling pathways linking calcium to these cellular functions are less understood. Calcium-dependent protein kinases (CDPKs), which are present in plants but not in animals, are also present in ciliated microorganisms which led us to examine their possible functions and mechanisms in flagellar related activities. By in silico analysis of Chlamydomonas genome we have identified 14 CDPKs and studied one of the flagellar localized CDPKs – CrCDPK3. CrCDPK3 was a protein of 485 amino acids and predicted to have a protein kinase domain at the N-terminus and four EF-hand motifs at the C-terminus. In flagella, CrCDPK3 was exclusively localized in the membrane matrix fraction and formed an unknown 20 S protein complex. Knockdown of CrCDPK3 expression by using artificial microRNA did not affect flagellar motility as well as flagellar adhesion and mating. Though flagellar shortening induced by treatment with sucrose or sodium pyrophosphate was not affected in RNAi strains, CrCDPK3 increased in the flagella, and pre-formed protein complex was disrupted. During flagellar regeneration, CrCDPK3 also increased in the flagella. When extracellular calcium was lowered to certain range by the addition of EGTA after deflagellation, flagellar regeneration was severely affected in RNAi cells compared with wild type cells. In addition, during flagellar elongation induced by LiCl, RNAi cells exhibited early onset of bulbed flagella. This work expands new functions of CDPKs in flagellar activities by showing involvement of CrCDPK3 in flagellar biogenesis in Chlamydomonas .

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Section: Resultssupporting

confidence: 88%

Section: Resultssupporting

confidence: 81%

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Regulation of Flagellar Biogenesis by a Calcium Dependent Protein Kinase in Chlamydomonas reinhardtii

Liang

Pan

2013

PLoS ONE

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“…Although LiCl induces flagellar growth (Nakamura et al , 1987; Periz et al , 2007), little growth occurs in the time during which IFT is recorded and 20 mM LiCl does not induce changes in the rate or frequency of anterograde or retrograde IFT (Dentler, 2005). Used cautiously, addition of 10–20 mM LiCl can facilitate recording IFT in motile flagella.…”

Section: Methodsmentioning

confidence: 99%

Recording and Analyzing IFT in Chlamydomonas Flagella

Dentler

VanderWaal

Porter

2009

Methods in Cell Biology

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The transport of materials to and from the cell body and tips of eukaryotic flagella and cilia is carried out by a process called intraflagellar transport, or IFT. This process is essential for the assembly and maintenance of cilia and flagella: in the absence of IFT, cilia cannot assemble and, if IFT is arrested in ciliated cells, the cilia disassemble. The major IFT complex proteins and the major motor proteins, kinesin-2 and osm-3 (which transport particles from the cell body to ciliary tips) and cytoplasmic dynein 1b (which transports particles from ciliary tips to the cell body) have been identified. However, we have little understanding of the structure of the IFT particles, the cargo that these particles carry, how cargo is loaded and unloaded from the particles, or how the motor proteins are regulated. The focus of this chapter is to provide methods to observe and quantify the movements of IFT particles in Chlamydomonas flagella. IFT movements can be visualized in paralyzed or partially arrested flagella using either differential interference contrast (IFT) microscopy or, in cells with fluorescently tagged IFT components, with fluorescence microscopy. Methods for recording IFT movements and analyzing movements using kymograms are described.

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“…Comparative genomic analysis (Avidor-Reiss et al, 2004;Chen et al, 2006;Li et al, 2004); transcriptional profiling (Blacque et al, 2005;Choksi et al, 2014;Periz et al, 2007;Stolc et al, 2005); and proteomic studies from Chlamydomonas flagella (Pazour et al, 2005), primary cilia (Ishikawa et al, 2012), motile cilia (Narita et al, 2012), and specialized mammalian cilia (Liu et al, 2007;Mayer et al, 2009) have identified numerous kinesin molecules. In addition, functional studies have discovered additional kinesins that are not identified in those large-scale analyses.…”

Section: Flagellar and Ciliary Kinesinsmentioning

confidence: 99%