Cruciform DNA binding protein in HeLa cell extracts

Biochemistry ◽  
1994 ◽  
Vol 33 (47) ◽  
pp. 14185-14196 ◽  
Author(s):  
Christopher E. Pearson ◽  
Marcia T. Ruiz ◽  
Gerald B. Price ◽  
Maria Zannis-Hadjopoulos
Keyword(s):  
Dna Binding ◽  
Hela Cell ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Cell Extracts ◽  
Cruciform Dna

Purification and properties of a cruciform DNA binding protein fromUstilago maydis

Chromosoma ◽  
1993 ◽  
Vol 102 (5) ◽  
pp. 348-354 ◽  
Author(s):  
H. Kotani ◽  
E. B. Kmiec ◽  
W. K. Holloman
Keyword(s):  
Dna Binding ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Purification And Properties ◽  
Cruciform Dna

scholarly journals The gene G13 in the class III region of the human MHC encodes a potential DNA-binding protein

1996 ◽  
Vol 319 (1) ◽  
pp. 81-89 ◽  
Author(s):  
Anu KHANNA ◽  
R. Duncan CAMPBELL
Keyword(s):  
Amino Acid ◽  
Dna Binding ◽  
Leucine Zipper ◽  
Binding Protein ◽  
Coiled Coil ◽  
Single Copy ◽  
Dna Binding Protein ◽  
Lymphoid Tissues ◽  
Class Iii ◽  
Cell Extracts

G13 is a single-copy gene lying approx. 75 kb centromeric of the complement gene cluster in the class III region of the human MHC. The gene spans approx. 17 kb of DNA and has been shown to encode mRNA of approx. 2.7 kb that is present in cell lines representing lymphoid and non-lymphoid tissues, indicating that it is ubiquitously expressed. The complete nucleotide sequence of the 2.7 kb mRNA has been derived from cDNA and genomic clones. The longest open reading frame obtained for G13 codes for a 703 amino acid protein of approx. 77 kDa in molecular mass. Comparison of the putative G13 amino acid sequence with the protein databases revealed significant similarities with DNA-binding proteins of the leucine zipper class, including a human cAMP response element binding protein. G13 contains a bZIP motif, a region rich in basic amino acids adjacent to a coiled-coil leucine zipper domain, common to this class of proteins that is known to be involved in dimerization and DNA binding. Antibodies raised against a fragment encoding the C-terminal half of the putative G13 protein recognized a major polypeptide of approx. 86 kDa and a minor polypeptide of approx. 78 kDa on immunoblotting of U937 cell extracts; this has been confirmed by immunoprecipitation experiments. Even though it contained at least one potential bipartite nuclear localization signal, the G13 protein was present both in the cytoplasm and the nucleus of the fibroblast cells. Thus G13 might be a novel DNA-binding protein that is perhaps translocated to the nucleus in a regulated manner.

scholarly journals CC1, a Novel Crenarchaeal DNA Binding Protein

2006 ◽  
Vol 189 (2) ◽  
pp. 403-409 ◽  
Author(s):  
Xiao Luo ◽  
Uli Schwarz-Linek ◽  
Catherine H. Botting ◽  
Reinhard Hensel ◽  
Bettina Siebers ◽  
...  
Keyword(s):  
Dna Binding ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Nucleic Acid Binding ◽  
Chromatin Protein ◽  
Gene Encoding ◽  
Single Stranded Dna ◽  
Cell Extracts ◽  
Domains Of Life ◽  
High Level

ABSTRACT The genomes of the related crenarchaea Pyrobaculum aerophilum and Thermoproteus tenax lack any obvious gene encoding a single-stranded DNA binding protein (SSB). SSBs are essential for DNA replication, recombination, and repair and are found in all other genomes across the three domains of life. These two archaeal genomes also have only one identifiable gene encoding a chromatin protein (the Alba protein), while most other archaea have at least two different abundant chromatin proteins. We performed a biochemical screen for novel nucleic acid binding proteins present in cell extracts of T. tenax. An assay for proteins capable of binding to a single-stranded DNA oligonucleotide resulted in identification of three proteins. The first protein, Alba, has been shown previously to bind single-stranded DNA as well as duplex DNA. The two other proteins, which we designated CC1 (for crenarchaeal chromatin protein 1), are very closely related to one another, and homologs are restricted to the P. aerophilum and Aeropyrum pernix genomes. CC1 is a 6-kDa, monomeric, basic protein that is expressed at a high level in T. tenax. This protein binds single- and double-stranded DNAs with similar affinities. These properties are consistent with a role for CC1 as a crenarchaeal chromatin protein.

scholarly journals Role of the Adenovirus DNA-Binding Protein in In Vitro Adeno-Associated Virus DNA Replication

1998 ◽  
Vol 72 (1) ◽  
pp. 420-427 ◽  
Author(s):  
Peter Ward ◽  
Frank B. Dean ◽  
Michael E. O’Donnell ◽  
Kenneth I. Berns
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Basic Question ◽  
Adeno Associated Virus ◽  
Vitro Assay ◽  
Cell Extracts ◽  
Vector Sequences

ABSTRACT A basic question in adeno-associated virus (AAV) biology has been whether adenovirus (Ad) infection provided any function which directly promoted replication of AAV DNA. Previously in vitro assays for AAV DNA replication, using linear duplex AAV DNA as the template, uninfected or Ad-infected HeLa cell extracts, and exogenous AAV Rep protein, demonstrated that Ad infection provides a direct helper effect for AAV DNA replication. It was shown that the nature of this helper effect was to increase the processivity of AAV DNA replication. Left unanswered was the question of whether this effect was the result of cellular factors whose activity was enhanced by Ad infection or was the result of direct participation of Ad proteins in AAV DNA replication. In this report, we show that in the in vitro assay, enhancement of processivity occurs with the addition of either the Ad DNA-binding protein (Ad-DBP) or the human single-stranded DNA-binding protein (replication protein A [RPA]). Clearly Ad-DBP is present after Ad infection but not before, whereas the cellular level of RPA is not apparently affected by Ad infection. However, we have not measured possible modifications of RPA which might occur after Ad infection and affect AAV DNA replication. When the substrate for replication was an AAV genome inserted into a plasmid vector, RPA was not an effective substitute for Ad-DBP. Extracts supplemented with Ad-DBP preferentially replicated AAV sequences rather than adjacent vector sequences; in contrast, extracts supplemented with RPA preferentially replicated vector sequences.

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