scholarly journals Alternative RNA Editing Produces a Novel Protein Involved in Mitochondrial DNA Maintenance in Trypanosomes

2008 ◽  
Vol 28 (18) ◽  
pp. 5595-5604 ◽  
Author(s):  
Torsten Ochsenreiter ◽  
Sedrick Anderson ◽  
Zachary A. Wood ◽  
Stephen L. Hajduk
Keyword(s):  
Dna Binding ◽  
Rna Editing ◽  
Basal Bodies ◽  
Kinetoplast Dna ◽  
Unusual Structure ◽  
Circular Dna ◽  
Modification Process ◽  
Dna Maintenance ◽  
First Time ◽  
Novel Protein

ABSTRACT The mitochondrial genome of trypanosomes is composed of thousands of topologically interlocked circular DNA molecules that form the kinetoplast DNA (kDNA). Most genes encoded by the kDNA require a posttranscriptional modification process called RNA editing to form functional mRNAs. Here, we show that alternative editing of the mitochondrial cytochrome c oxidase III (COXIII) mRNA in Trypanosoma brucei produces a novel DNA binding protein, alternatively edited protein 1 (AEP-1). AEP-1 localizes to the region of the cell between the kDNA and the flagellum and purifies with the tripartite attachment complex, a structure believed to physically link the kDNA and flagellar basal bodies. Expression of the DNA binding domain of AEP-1 results in aberrant kDNA structure and reduced cell growth, indicating that AEP-1 is involved in the maintenance of the kDNA. Perhaps most important, our studies show a gain of function through an alternatively edited mRNA and, for the first time, provide a link between the unusual structure of the kDNA and RNA editing in trypanosome mitochondria.

scholarly journals DisAp-dependent striated fiber elongation is required to organize ciliary arrays

2014 ◽  
Vol 207 (6) ◽  
pp. 705-715 ◽  
Author(s):  
Domenico F. Galati ◽  
Stephanie Bonney ◽  
Zev Kronenberg ◽  
Christina Clarissa ◽  
Mark Yandell ◽  
...  
Keyword(s):  
Basal Bodies ◽  
The Novel ◽  
Fiber Elongation ◽  
Ciliary Beating ◽  
Attachment Sites ◽  
Multiciliated Cells ◽  
Fiber Protein ◽  
Striated Fiber ◽  
First Time ◽  
Novel Protein

Cilia-organizing basal bodies (BBs) are microtubule scaffolds that are visibly asymmetrical because they have attached auxiliary structures, such as striated fibers. In multiciliated cells, BB orientation aligns to ensure coherent ciliary beating, but the mechanisms that maintain BB orientation are unclear. For the first time in Tetrahymena thermophila, we use comparative whole-genome sequencing to identify the mutation in the BB disorientation mutant disA-1. disA-1 abolishes the localization of the novel protein DisAp to T. thermophila striated fibers (kinetodesmal fibers; KFs), which is consistent with DisAp’s similarity to the striated fiber protein SF-assemblin. We demonstrate that DisAp is required for KFs to elongate and to resist BB disorientation in response to ciliary forces. Newly formed BBs move along KFs as they approach their cortical attachment sites. However, because they contain short KFs that are rotated, BBs in disA-1 cells display aberrant spacing and disorientation. Therefore, DisAp is a novel KF component that is essential for force-dependent KF elongation and BB orientation in multiciliary arrays.

Cloning and characterization of E2F-2, a novel protein with the biochemical properties of transcription factor E2F

1993 ◽  
Vol 13 (12) ◽  
pp. 7802-7812
Author(s):  
M Ivey-Hoyle ◽  
R Conroy ◽  
H E Huber ◽  
P J Goodhart ◽  
A Oliff ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Dna Binding ◽  
Hela Cell ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Amino Acid Identity ◽  
Biochemical Properties ◽  
Binding Activity ◽  
Dna Binding Activity ◽  
Novel Protein

E2F is a mammalian transcription factor that appears to play an important role in cell cycle regulation. While at least two proteins (E2F-1 and DP-1) with E2F-like activity have been cloned, studies from several laboratories suggest that additional homologs may exist. A novel protein with E2F-like properties, designated E2F-2, was cloned by screening a HeLa cDNA library with a DNA probe derived from the DNA binding domain of E2F-1 (K. Helin, J. A. Lees, M. Vidal, N. Dyson, E. Harlow, and A. Fattaey, Cell 70:337-350, 1992). E2F-2 exhibits overall 46% amino acid identity to E2F-1. Both the sequence and the function of the DNA and retinoblastoma gene product binding domains of E2F-1 are conserved in E2F-2. The DNA binding activity of E2F-2 is dramatically enhanced by complementation with particular sodium dodecyl sulfate-polyacrylamide gel electrophoresis-purified components of HeLa cell E2F, and anti-E2F-2 antibodies cross-react with components of purified HeLa cell E2F. These observations are consistent with a model in which E2F binds DNA as a heterodimer of two distinct proteins, and E2F-2 is functionally and immunologically related to one of these proteins.

A 210 kDa protein is located in a membrane-microtubule linker at the distal end of mature and nascent basal bodies

1999 ◽  
Vol 112 (11) ◽  
pp. 1633-1644 ◽  
Author(s):  
K.F. Lechtreck ◽  
A. Teltenkotter ◽  
A. Grunow
Keyword(s):  
Basal Body ◽  
Body Position ◽  
Immunogold Electron Microscopy ◽  
Basal Bodies ◽  
Western Blots ◽  
Whole Cells ◽  
Extracellular Matrix Components ◽  
Green Flagellate ◽  
Protein Rich ◽  
Novel Protein

A monoclonal antibody raised against purified flagellar basal apparatuses from the green flagellate Spermatozopsis similis reacted with a protein of 210 kDa (p210) in western blots. The protein was partially cloned by immunoscreening of a cDNA library. The sequence encoded a novel protein rich in alanine (25%) and proline (20%), which contained regions similar to proteins of comparable amino acid composition such as extracellular matrix components or the membrane-cytoskeletal linker synapsin. Using a polyclonal antibody (anti-p210) raised against the C-terminal part of p210, it was shown that the protein was highly enriched in the basal apparatuses. Immunogold electron microscopy of isolated cytoskeletons or whole cells revealed that p210 was located in the flagellar transition region. The protein was part of the Y-shaped fibrous linkers between the doublet microtubules and the flagellar membrane, as indicated by statistical analysis of post-labeled sections using anti-centrin and anti-tubulin as controls. In premitotic cells p210 was located in a fibrous layer at the distal end of nascent basal bodies, which was perforated by the outgrowing axoneme. During deflagellation the protein remained at the basal body but we observed changes in its distribution, indicating that p210 partially moved to the tip of the basal body. p210 can be used as a marker to determine basal body position, orientation (parallel or antiparallel) and number in S. similis by indirect immunofluorescence. We suppose that p210 is involved in linking basal bodies to the plasma membrane, which is an important step during ciliogenesis.

Photodegradable self-assembling PAMAM dendrons for gene delivery involving dendriplex formation and phototriggered circular DNA release

2016 ◽  
Vol 40 (3) ◽  
pp. 2601-2608 ◽  
Author(s):  
Yu-Sen Lai ◽  
Chai-Lin Kao ◽  
Ya-Pei Chen ◽  
Chia-Chia Fang ◽  
Chao-Chin Hu ◽  
...  
Keyword(s):  
Dna Binding ◽  
Gene Delivery ◽  
Self Assembly ◽  
Circular Dna ◽  
Self Assembling ◽  
Dna Release

Photoresponsive amphiphilic dendron bearing a photolabile o-nitrobenzyl group possesses self-assembly, DNA binding and photo-induced release.

scholarly journals ClpX is Essential and Activated by Single Strand DNA Binding Protein in Mycobacteria

2020 ◽  
Author(s):  
Jemila C. Kester ◽  
Olga Kandror ◽  
Tatos Akopian ◽  
Michael R. Chase ◽  
Junhao Zhu ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Dna Replication ◽  
Dna Binding ◽  
Drug Targets ◽  
Atp Hydrolysis ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Cellular Functions ◽  
Mycobacterial Growth ◽  
Dna Maintenance

The ClpP1P2 proteolytic complex is essential in Mycobacterium tuberculosis (Mtb). Proteolysis by ClpP1P2 requires an associated ATPase, either ClpX or ClpC1. Here, we seek to define the unique contributions of the ClpX ATPase to mycobacterial growth. We formally demonstrate that ClpX is essential for mycobacterial growth and to understand its essential functions, we identify ClpX-His-interacting proteins by pulldown and tandem mass spectrometry. We find an unexpected association between ClpX and proteins involved in DNA replication, and confirm a physical association between ClpX and the essential DNA maintenance protein Single-Stranded DNA Binding protein (SSB). Purified SSB is not degraded by ClpXP1P2; instead SSB enhances ATP hydrolysis by ClpX and degradation of the model substrate GFP-SsrA by ClpXP1P2. This activation of ClpX is mediated by the C-terminal tail of SSB that had been implicated in the activation of other ATPases associated with DNA replication. Consistent with the predicted interactions, depletion of clpX transcript perturbs DNA replication. These data reveal that ClpX participates in DNA replication and identify the first activator of ClpX in mycobacteria. IMPORTANCE Tuberculosis, caused by Mycobacterium tuberculosis, imposes a major global health burden, surpassing HIV and malaria in annual deaths. The ClpP1P2 proteolytic complex and its cofactor ClpX are attractive drug targets, but their precise cellular functions are unclear. This work confirms ClpX’s essentiality and describes a novel interaction between ClpX and SSB, a component of the DNA replication machinery. Further, we demonstrate that a loss of ClpX is sufficient to interrupt DNA replication, suggesting the ClpX-SSB complex may play a role in DNA replication in mycobacteria.

An electrochemical sensing platform based on local repression of electrolyte diffusion for single-step, reagentless, sensitive detection of a sequence-specific DNA-binding protein

The Analyst ◽  
2014 ◽  
Vol 139 (9) ◽  
pp. 2193-2198 ◽  
Author(s):  
Yun Zhang ◽  
Fang Liu ◽  
Jinfang Nie ◽  
Fuyang Jiang ◽  
Caibin Zhou ◽  
...  
Keyword(s):  
Dna Binding ◽  
Electrode Surface ◽  
Electrochemical Biosensor ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Single Step ◽  
Dna Probe ◽  
Electrochemical Sensing ◽  
Sensing Platform ◽  
First Time

This paper describes for the first time an electrochemical biosensor, which employs a DNA probe modified with a redox tag close to electrode surface, for picomolar detection of a sequence-specific DNA-binding protein.

scholarly journals A novel protein with DNA binding activity from tobacco chloroplast nucleoids.

1997 ◽  
Vol 9 (9) ◽  
pp. 1673-1682 ◽  
Author(s):  
T Nakano ◽  
S Murakami ◽  
T Shoji ◽  
S Yoshida ◽  
Y Yamada ◽  
...  
Keyword(s):  
Dna Binding ◽  
Binding Activity ◽  
Tobacco Chloroplast ◽  
Dna Binding Activity ◽  
Chloroplast Nucleoids ◽  
Novel Protein

Taxodione, a DNA-Binding Compound from Taxodium distichum L. (Rich.)

2008 ◽  
Vol 63 (5-6) ◽  
pp. 355-360 ◽  
Author(s):  
Ahmed M. Zaghloul ◽  
Ahmed A. Gohar ◽  
Zein Al-Abdin M. Naiem ◽  
Fatma M. Abdel Bar
Keyword(s):  
Dna Binding ◽  
Hepatic Stellate Cells ◽  
Shikimic Acid ◽  
Antitumour Activity ◽  
Stellate Cells ◽  
Taxodium Distichum ◽  
Affinity Probe ◽  
First Time

8-β-Hydroxypimar-15-en-19-oic acid (1), taxodione (2), 6,7-dehydro-8-hydrotaxodone (3), quercetin-3-O-β-d-glucopyranoside (4), and shikimic acid (5) were isolated from the leaves of Taxodium distichum L. (Rich.) for the first time. Previously reported compounds [β-sitosterol (6), isorhamnetin (7), quercetin (8), isorhamnetin-3-O-α-arabinofuranoside (9), quercetin- 3-O-α-arabinofuranoside (10)] have also been isolated. The activity of taxodione as an inhibitor for hepatic stellate cells was determined. The antitumour activity of 2, 3, and 5 using a DNA affinity probe was examined.

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