Changes in protein and gene expression during induction of the CO2-concentrating mechanism in wild-type and mutant Chlamydomonas

1991 ◽  
Vol 69 (5) ◽  
pp. 1008-1016 ◽  
Author(s):  
Martin H. Spalding ◽  
Thomas L. Winder ◽  
James C. Anderson ◽  
Anne M. Geraghty ◽  
Laura F. Marek

Several changes occur in wild-type Chlamydomonas reinhardtii upon exposure to limiting CO2, including induction of several polypeptides. Polypeptide induction was previously shown to correlate with appearance of the active CO2-concentrating mechanism (CCM) of this alga. In this paper induction of polypeptides by limiting CO2 was investigated in mutants with lesions in the CCM. Mutants with lesions in the ca-1 and pmp-1 loci exhibited alterations in polypeptide induction, but it was concluded that the alterations probably do not represent their primary genetic lesions. Other changes that occur in this alga in response to limiting CO2 were also investigated. Based on a lack of significant change in the transcript abundance of ribulose-1,5-bisphosphate carboxylase/oxygenase large and small subunit genes in the wild type, it was concluded that the previously reported transient decline in synthesis of both subunits is controlled at the translational level. A transient increase in the activity of the photorespiratory enzyme phosphoglycolate phosphatase was observed in the wild type but not in a mutant, cia-5, that lacks induction of the CCM. In addition, changes in expression of genes encoding periplasmic carbonic anhydrase, a 36-kDa membrane-associated protein and a chlorophyll-binding protein occurred in the wild type but not in cia-5 in response to limiting CO2. The absence of these changes in cia-5 was attributed to a lack of either the signal itself or transduction of the signal responsible for adaptation to limiting CO2, which led to speculation that a larger range of responses are regulated by the same signal than was previously recognized. Key words: photosynthesis, photorespiration, algae, inorganic carbon transport, transcription, translation.

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Koumei Yazaki ◽  
Chinatsu Yoshikoshi ◽  
Satoru Oshiro ◽  
Sumino Yanase

Astaxanthin (AX), which is produced by some marine animals, is a type of carotenoid that has antioxidative properties. In this study, we initially examined the effects of AX on the aging of a model organismC. elegansthat has the conserved intracellular pathways related to mammalian longevity. The continuous treatments with AX (0.1 to 1 mM) from both the prereproductive and young adult stages extended the mean lifespans by about 16–30% in the wild-type and long-lived mutantage-1ofC. elegans. In contrast, the AX-dependent lifespan extension was not observed even in adaf-16null mutant. Especially, the expression of genes encoding superoxide dismutases and catalases increased in two weeks after hatching, and the DAF-16 protein was translocated to the nucleus in the AX-exposed wild type. These results suggest that AX protects the cell organelle mitochondria and nucleus of the nematode, resulting in a lifespan extension via an Ins/IGF-1 signaling pathway during normal aging, at least in part.


1998 ◽  
Vol 76 (6) ◽  
pp. 1035-1042 ◽  
Author(s):  
Hiroshi Ohkawa ◽  
Masatoshi Sonoda ◽  
Hirokazu Katoh ◽  
Teruo Ogawa

Mutants of cyanobacteria defective in parts of the CO2-concentrating mechanism are classified into three types. (i) Mutants defective in inorganic carbon transporters. One of these mutants was constructed by inactivating cmpA encoding 42 kDa protein in the cytoplasmic membrane. (ii) Mutants defective in NAD(P)H dehydrogenase(s). There are five ndhD genes in Synechocystis PCC6803, two of them expressed constitutively and three inducible by low CO2. Two kinds of NAD(P)H dehydrogenase appear to be involved in energizing and inducing the high affinity inorganic carbon transport system. (iii) Mutants defective in carboxysome with impaired ccm or icfA genes. New type of mutants with impaired cotA (renamed as pxcA) have also been isolated. These mutants did not show light-induced proton extrusion and were unable to grow at acidic pHs. A mutant constructed by inactivating cotA (pxcA) in the wild-type Synechocystis was unable to transport CO2 at pH 6.5. We concluded that cotA (pxcA) has a role in light-induced proton extrusion that is essential at acidic pHs to extrude protons produced during CO2 transport.Key words: CO2-concentrating mechanism (CCM), CO2 transport, NAD(P)H dehydrogenase, proton extrusion, carboxysome, mutant.


Development ◽  
1993 ◽  
Vol 119 (Supplement) ◽  
pp. 149-161 ◽  
Author(s):  
Michael Bate ◽  
Emma Rushton ◽  
Manfred Frasch

In wild-type embryos of Drosophila melanogaster, the formation of differentiated larval muscles is preceded by the segregation of small numbers of progenitor or founder cells in the embryonic mesoderm. The founder cells, characterised by the expression of genes encoding putative transcription factors such as S59 or vestigial, fuse with neighbouring myoblasts to form syncytial pre­ cursors of individual muscles. Founder cell segregation is deranged in embryos mutant for any of the neurogenic genes: enlarged clusters of cells expressing S59 or vestigial are detected at the sites where small numbers of founder cells segregate in the wild type. In addition, muscle differentiation is deranged in such embryos in a way that appears to be closely linked to the extent of epidermal disruption caused by the neurogenic phenotype: myoblast fusion is limited to regions of the mesoderm beneath the residual epidermis left by the hyperplasia of the nervous system, and late expression of S59 and vestigial is lost from mesoderm not lying within the margins of the residual epidermis. Thus neurogenic gene functions appear to be required both for the normal segregation of founder cells and for muscle differentiation. It is not clear whether either of these requirements reflects an essential function for any or all of the neurogenic genes within the mesoderm itself.


1991 ◽  
Vol 69 (5) ◽  
pp. 945-950 ◽  
Author(s):  
Judy Lieman-Hurwitz ◽  
Rakefet Schwarz ◽  
Flor Martinez ◽  
Zeev Maor ◽  
Leonora Reinhold ◽  
...  

Modifications of the genomic region near (and including) rbc, the operon that codes for the large and small subunits of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), resulted in cyanobacterial mutants that demand high CO2 for growth. Mutant EK6 originated from the fusion of the 3' end of rbcS (which codes for the small subunit of Rubisco) with 84 nucleotides from the 5′ flanking region of nptII (kanamycin-resistance gene), leading to a 17-kDa small subunit, as compared to 14 kDa in the wild type. Mutant D4 originated from substitution of the 1.4-kb PstI fragment, downstream of rbc, with nptII, inactivating several open reading frames in this region. Mutant O105 was obtained by chemical mutagenesis and the mutation was mapped approximately 9 kb upstream of rbc. Mutants EK6 and O105 exhibited a very low apparent photosynthetic affinity for inorganic carbon, whereas D4 had an affinity similar to that observed in wild-type cells grown under high CO2. These mutants, and the constructs used to raise them, can be used to study the role of the small subunit of Rubisco and the genomic region near rbc in cyanobacterial photosynthesis. We propose that this region contains a cluster of genes involved in the ability of cyanobacteria to grow under low ambient CO2..


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
José Francisco Cruz-Pérez ◽  
Roxana Lara-Oueilhe ◽  
Cynthia Marcos-Jiménez ◽  
Ricardo Cuatlayotl-Olarte ◽  
María Luisa Xiqui-Vázquez ◽  
...  

AbstractThe plant growth-promoting bacterium Azospirillum brasilense contains several genes encoding proteins involved in the biosynthesis and degradation of the second messenger cyclic-di-GMP, which may control key bacterial functions, such as biofilm formation and motility. Here, we analysed the function and expression of the cdgD gene, encoding a multidomain protein that includes GGDEF-EAL domains and CHASE and PAS domains. An insertional cdgD gene mutant was constructed, and analysis of biofilm and extracellular polymeric substance production, as well as the motility phenotype indicated that cdgD encoded a functional diguanylate protein. These results were correlated with a reduced overall cellular concentration of cyclic-di-GMP in the mutant over 48 h compared with that observed in the wild-type strain, which was recovered in the complemented strain. In addition, cdgD gene expression was measured in cells growing under planktonic or biofilm conditions, and differential expression was observed when KNO3 or NH4Cl was added to the minimal medium as a nitrogen source. The transcriptional fusion of the cdgD promoter with the gene encoding the autofluorescent mCherry protein indicated that the cdgD gene was expressed both under abiotic conditions and in association with wheat roots. Reduced colonization of wheat roots was observed for the mutant compared with the wild-type strain grown in the same soil conditions. The Azospirillum-plant association begins with the motility of the bacterium towards the plant rhizosphere followed by the adsorption and adherence of these bacteria to plant roots. Therefore, it is important to study the genes that contribute to this initial interaction of the bacterium with its host plant.


2020 ◽  
Author(s):  
Maik Hintze ◽  
Sebastian Griesing ◽  
Marion Michels ◽  
Birgit Blanck ◽  
Lena Wischhof ◽  
...  

AbstractWe investigated the contribution of apoptosis-inducing factor (AIF), a key regulator of mitochondrial biogenesis, in supporting hair growth. We report that pelage abnormalities developed during hair follicle (HF) morphogenesis in Harlequin (Hq) mutant mice. Fragility of the hair cortex was associated with decreased expression of genes encoding structural hair proteins, though key transcriptional regulators of HF development were expressed at normal levels. Notably, Aifm1 (R200 del) knockin males and Aifm1(R200 del)/Hq females showed minor hair defects, despite substantially reduced AIF levels. Furthermore, we cloned the integrated ecotropic provirus of the Aifm1Hq allele. We found that its overexpression in wild-type keratinocyte cell lines led to down-regulation of HF-specific Krt84 and Krtap3-3 genes without altering Aifm1 or epidermal Krt5 expression. Together, our findings imply that pelage paucity in Hq mutant mice is mechanistically linked to severe AIF deficiency and is associated with the expression of retroviral elements that might potentially influence the transcriptional regulation of structural hair proteins.


Microbiology ◽  
2006 ◽  
Vol 152 (7) ◽  
pp. 2003-2012 ◽  
Author(s):  
Verena Seidl ◽  
Irina S. Druzhinina ◽  
Christian P. Kubicek

To identify carbon sources that trigger β-N-acetylglucosaminidase (NAGase) formation in Hypocrea atroviridis (anamorph Trichoderma atroviride), a screening system was designed that consists of a combination of Biolog Phenotype MicroArray plates, which contain 95 different carbon sources, and specific enzyme activity measurements using a chromogenic substrate. The results revealed growth-dependent kinetics of NAGase formation and it was shown that NAGase activities were enhanced on carbon sources sharing certain structural properties, especially on α-glucans (e.g. glycogen, dextrin and maltotriose) and oligosaccharides containing galactose. Enzyme activities were assessed in the wild-type and a H. atroviridis Δnag1 strain to investigate the influence of the two NAGases, Nag1 and Nag2, on total NAGase activity. Reduction of NAGase levels in the Δnag1 strain in comparison to the wild-type was strongly carbon-source and growth-phase dependent, indicating the distinct physiological roles of the two proteins. The transcript abundance of nag1 and nag2 was increased on carbon sources with elevated NAGase activity, indicating transcriptional regulation of these genes. The screening method for the identification of carbon sources that induce enzymes or a gene of interest, as presented in this paper, can be adapted for other purposes if appropriate enzyme or reporter assays are available.


Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 17 ◽  
Author(s):  
Guang Chen ◽  
Jiang Hu ◽  
Liuliu Dong ◽  
Dali Zeng ◽  
Longbiao Guo ◽  
...  

A panel of ethane-methyl-sulfonate-mutagenized japonica rice lines was grown in the presence of salinity in order to identify genes required for the expression of salinity tolerance. A highly nontolerant selection proved to harbor a mutation in FLN2, a gene which encodes fructokinase-like protein2. Exposure of wild-type rice to salinity up-regulated FLN2, while a CRISPR/Cas9-generated FLN2 knockout line was hypersensitive to the stress. Both ribulose 1,5-bisphosphate carboxylase/oxygenase activity and the abundance of the transcript generated by a number of genes encoding components of sucrose synthesis were lower in the knockout line than in wild-type plants’ leaves, while the sucrose contents of the leaf and root were, respectively, markedly increased and decreased. That sugar partitioning to the roots was impaired in FLN2 knockout plants was confirmed by the observation that several genes involved in carbon transport were down-regulated in both the leaf and in the leaf sheath. The levels of sucrose synthase, acid invertase, and neutral invertase activity were distinctly lower in the knockout plants’ roots than in those of wild-type plants, particularly when the plants were exposed to salinity stress. The compromised salinity tolerance exhibited by the FLN2 knockout plants was likely a consequence of an inadequate supply of the assimilate required to support growth, a problem which was rectifiable by providing an exogenous supply of sucrose. The conclusion was that FLN2, on account of its influence over sugar metabolism, is important in the context of seedling growth and the rice plant’s response to salinity stress.


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