scholarly journals Role of Two NtcA-Binding Sites in the Complex ntcA Gene Promoter of the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120

2008 ◽  
Vol 190 (22) ◽  
pp. 7584-7590 ◽  
Author(s):  
Elvira Olmedo-Verd ◽  
Ana Valladares ◽  
Enrique Flores ◽  
Antonia Herrero ◽  
Alicia M. Muro-Pastor
Keyword(s):  
Binding Sites ◽  
Gene Promoter ◽  
Filamentous Cyanobacterium ◽  
Mobility Shift ◽  
Dnase I Footprinting ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120

ABSTRACT Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that fixes N2 in specialized cells called heterocysts, which differentiate from vegetative cells in a process that requires the nitrogen control transcription factor NtcA. 2-Oxoglutarate-stimulated binding of purified NtcA to wild-type and modified versions of the ntcA gene promoter from Anabaena sp. was analyzed by mobility shift and DNase I footprinting assays, and the role of NtcA-binding sites in the expression of the ntcA gene during heterocyst differentiation was studied in vivo by using an ntcA-gfp translational fusion and primer extension analysis. Mutation of neither of the two identified NtcA-binding sites eliminated localized expression of ntcA in proheterocysts, but mutation of both sites led to very low, nonlocalized expression.

scholarly journals Manipulation of Pattern of Cell Differentiation in a hetR Mutant of Anabaena sp. PCC 7120 by Overexpressing hetZ Alone or with hetP

Life ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 60 ◽  
Author(s):  
He Zhang ◽  
Xudong Xu
Keyword(s):  
Cell Differentiation ◽  
Peptide Inhibitor ◽  
Filamentous Cyanobacterium ◽  
Heterocyst Differentiation ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120

In the filamentous cyanobacterium, Anabaena sp. PCC 7120, single heterocysts differentiate at semi-regular intervals in response to nitrogen stepdown. HetR is a principal regulator of heterocyst differentiation, and hetP and hetZ are two genes that are regulated directly by HetR. In a hetR mutant generated from the IHB (Institute of Hydrobiology) substrain of PCC 7120, heterocyst formation can be restored by moderate expression of hetZ and hetP. The resulting heterocysts are located at terminal positions. We used a tandem promoter, PrbcLPpetE, to express hetZ and hetP strongly in the hetR mutant. Co-expression of hetZ and hetP enabled the hetR mutant to form multiple contiguous heterocysts at both terminal and intercalary positions. Expression of hetZ, alone resulted in terminally located heterocysts, whereas expression of hetP, alone produced enlarged cells in strings. In the absence of HetR, formation of heterocysts was insensitive to the peptide inhibitor, RGSGR.

scholarly journals Inactivation of a Heterocyst-Specific Invertase Indicates a Principal Role of Sucrose Catabolism in Heterocysts of Anabaena sp.

2010 ◽  
Vol 192 (20) ◽  
pp. 5526-5533 ◽  
Author(s):  
Rocío López-Igual ◽  
Enrique Flores ◽  
Antonia Herrero
Keyword(s):  
Fluorescent Protein ◽  
Northern Analysis ◽  
Filamentous Cyanobacterium ◽  
Nitrogen Deprivation ◽  
Vegetative Cells ◽  
Anabaena Sp ◽  
Green Fluorescent ◽  
Pcc 7120 ◽  
Diazotrophic Growth

ABSTRACT Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that carries out N2 fixation in specialized cells called heterocysts, which exchange nutrients and regulators with the filament's vegetative cells that perform the photosynthetic fixation of CO2. The Anabaena genome carries two genes coding for alkaline/neutral invertases, invA and invB. As shown by Northern analysis, both genes were expressed monocistronically and induced under nitrogen deprivation, although induction was stronger for invB than for invA. Whereas expression of an InvA-N-GFP fusion (green fluorescent protein [GFP] fused to the N terminus of the InvA protein [InvA-N]) was homogeneous along the cyanobacterial filament, consistent with the lack of dependence on HetR, expression of an InvB-N-GFP fusion upon combined nitrogen deprivation took place mainly in differentiating and mature heterocysts. In an hetR genetic background, the InvB-N-GFP fusion was strongly expressed all along the filament. An insertional mutant of invA could grow diazotrophically but was impaired in nifHDK induction and exhibited an increased frequency of heterocysts, suggesting a regulatory role of the invertase-mediated carbon flux in vegetative cells. In contrast, an invB mutant was strongly impaired in diazotrophic growth, showing a crucial role of sucrose catabolism mediated by the InvB invertase in the heterocysts.

Distinct functional contributions of 2 GABP–NRF-2 recognition sites within the context of the human TOMM70 promoter

2006 ◽  
Vol 84 (5) ◽  
pp. 813-822 ◽  
Author(s):  
José R. Blesa ◽  
José Hernández-Yago
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Binding Sites ◽  
Site Directed Mutagenesis ◽  
Outer Mitochondrial Membrane ◽  
Mobility Shift ◽  
A Subunit ◽  
Expression Evidence ◽  
Electrophoretic Mobility Shift Assays

TOMM70 is a subunit of the outer mitochondrial membrane translocase that plays a major role as a receptor of hydrophobic preproteins targeted to mitochondria. We have previously reported 2 binding sites for the transcription factor GABP–NRF-2 in the promoter region of the human TOMM70 gene that are important in activating transcription. To assess the functionality and actual role of these sites, chromatin immunoprecipitation, site-directed mutagenesis, and electrophoretic mobility shift assays were carried out. We conclude that GABP–NRF-2 binds in vivo to the TOMM70 promoter, and that the 2 GABP–NRF-2 binding sites of the promoter have different functional contributions in promoting TOMM70 expression. Evidence is provided that they work in an additive manner as single sites.

scholarly journals Transcriptional role of a conserved GATA-1 site in the human epsilon-globin gene promoter.

1991 ◽  
Vol 11 (5) ◽  
pp. 2558-2566 ◽  
Author(s):  
Q H Gong ◽  
J Stern ◽  
A Dean
Keyword(s):  
Globin Gene ◽  
Point Mutations ◽  
Gene Promoter ◽  
Globin Genes ◽  
Mobility Shift ◽  
Dnase I Footprinting ◽  
Nuclear Extracts ◽  
Protein Dna Interactions ◽  
Globin Promoter

The epsilon-globin gene is the first of the human beta-like globin genes to be expressed during development. We have analyzed protein-DNA interactions in the epsilon-globin promoter region by DNase I footprinting and electrophoretic mobility shift experiments using nuclear extracts from K562 human erythroid cells and from nonerythroid HeLa cells. A restricted set of ubiquitous proteins, including Sp1, bound to regions of the promoter including the CACCC and CCAAT sites. Three interactions, at positions -213, -165, and +3 relative to the transcription start site, were erythroid specific and corresponded to binding of GATA-1, a transcription factor highly restricted to the erythroid lineage. Interestingly, the GATA-1 site at -165 has been conserved in the promoters of 10 mammalian embryonic globin genes. Point mutations demonstrate that GATA-1 binding to this site is necessary for interaction with an erythroid-specific enhancer but that in the absence of an enhancer, GATA-1 does not increase transcription.

Early candidacy for differentiation into heterocysts in the filamentous cyanobacterium Anabaena sp. PCC 7120

2009 ◽  
Vol 192 (1) ◽  
pp. 23-31 ◽  
Author(s):  
Masakazu Toyoshima ◽  
Naobumi V. Sasaki ◽  
Makoto Fujiwara ◽  
Shigeki Ehira ◽  
Masayuki Ohmori ◽  
...  
Keyword(s):  
Filamentous Cyanobacterium ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120

An increase in the level of 2-oxoglutarate promotes heterocyst development in the cyanobacterium Anabaena sp. strain PCC 7120

Microbiology ◽  
2003 ◽  
Vol 149 (11) ◽  
pp. 3257-3263 ◽  
Author(s):  
Jian-Hong Li ◽  
Sophie Laurent ◽  
Viren Konde ◽  
Sylvie Bédu ◽  
Cheng-Cai Zhang
Keyword(s):  
Inorganic Nitrogen ◽  
Cell Types ◽  
Filamentous Cyanobacterium ◽  
Nitrogen Status ◽  
Gene Encoding ◽  
Combined Nitrogen ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120 ◽  
Heterocyst Development

In the filamentous cyanobacterium Anabaena sp. strain PCC 7120, a starvation of combined nitrogen induces differentiation of heterocysts, cells specialized in nitrogen fixation. How do filaments perceive the limitation of the source of combined nitrogen, and what determines the proportion of heterocysts? In cyanobacteria, 2-oxoglutarate provides a carbon skeleton for the incorporation of inorganic nitrogen. Recently, it has been proposed that the concentration of 2-oxoglutarate reflects the nitrogen status in cyanobacteria. To investigate the effect of 2-oxoglutarate on heterocyst development, a heterologous gene encoding a 2-oxoglutarate permease under the control of a regulated promoter was expressed in Anabaena sp. PCC 7120. The increase of 2-oxoglutarate within cells can trigger heterocyst differentiation in a subpopulation of filaments even in the presence of nitrate. In the absence of a source of combined nitrogen, it can increase heterocyst frequency, advance the timing of commitment to heterocyst development and further increase the proportion of heterocysts in a patS mutant. Here, it is proposed that the intracellular concentration of 2-oxoglutarate is involved in the determination of the proportion of the two cell types according to the carbon/nitrogen status of the filament.

Function analysis of RNase E in the filamentous cyanobacterium Anabaena sp. PCC 7120

2020 ◽  
Vol 171 (5-6) ◽  
pp. 194-202
Author(s):  
Huaduo Yan ◽  
Yarui Cheng ◽  
Li Wang ◽  
Wenli Chen
Keyword(s):  
Function Analysis ◽  
Filamentous Cyanobacterium ◽  
Rnase E ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120

Characterization and responses to environmental cues of a photosynthetic antenna-deficient mutant of the filamentous cyanobacterium Anabaena sp. PCC 7120

2014 ◽  
Vol 171 (11) ◽  
pp. 915-926 ◽  
Author(s):  
Francisco Leganés ◽  
Francisco Martínez-Granero ◽  
M. Ángeles Muñoz-Martín ◽  
Eduardo Marco ◽  
Alberto Jorge ◽  
...  
Keyword(s):  
Environmental Cues ◽  
Filamentous Cyanobacterium ◽  
Deficient Mutant ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120 ◽  
Photosynthetic Antenna

scholarly journals In Vitro and In Vivo Analysis of the Role of PrrA in Rhodobacter sphaeroides 2.4.1 hemA Gene Expression

2006 ◽  
Vol 188 (9) ◽  
pp. 3208-3218 ◽  
Author(s):  
Britton Ranson-Olson ◽  
Denise F. Jones ◽  
Timothy J. Donohue ◽  
Jill H. Zeilstra-Ryalls
Keyword(s):  
Rhodobacter Sphaeroides ◽  
Dna Sequences ◽  
Binding Sites ◽  
Aminolevulinic Acid ◽  
Regulation Of Transcription ◽  
Mobility Shift ◽  
Metabolic Switch

ABSTRACT The hemA gene codes for one of two synthases in Rhodobacter sphaeroides 2.4.1 which catalyze the formation of 5-aminolevulinic acid. We have examined the role of PrrA, a DNA binding protein that is associated with the metabolic switch between aerobic growth and anoxygenic photosynthetic growth, in hemA expression and found that hemA transcription is directly activated by PrrA. Using electrophoretic mobility shift assays and DNase I protection assays, we have mapped two binding sites for PrrA within the hemA upstream sequences, each of which contains an identical 9-bp motif. Using lacZ transcription reporter plasmids in wild-type strain 2.4.1 and PrrA− mutant strain PRRA2, we showed that PrrA was required for maximal expression. We also found that the relative impacts of altering DNA sequences within the two binding sites are different depending on whether cells are growing aerobically or anaerobically. This reveals a greater level of complexity associated with PrrA-mediated regulation of transcription than has been heretofore described. Our findings are of particular importance with respect to those genes regulated by PrrA having more than one upstream binding site. In the case of the hemA gene, we discuss possibilities as to how these new insights can be accommodated within the context of what has already been established for hemA transcription regulation in R. sphaeroides.

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