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Article: Genome-wide analysis for the melatonin trait associated genes and SNPs in dairy goat (

Wu, Hao / Yi, Qi / Ma, Wenkui / Yan, Laiqing / Guan, Shengyu / Wang, Likai / Yang, Guang / Tan, Xinxing / Ji, Pengyun / Liu, Guoshi

Frontiers in genetics

2023 Volume 14, Page(s) 1118367

Abstract: Previous studies have reported that the endogenous melatonin level is positively associated with the quality and yield of milk of cows. In the current study, a total of 34,921 SNPs involving 1,177 genes were identified in dairy goats by using the whole ... ...

Abstract	Previous studies have reported that the endogenous melatonin level is positively associated with the quality and yield of milk of cows. In the current study, a total of 34,921 SNPs involving 1,177 genes were identified in dairy goats by using the whole genome resequencing bulked segregant analysis (BSA) analysis. These SNPs have been used to match the melatonin levels of the dairy goats. Among them, 3 SNPs has been identified to significantly correlate with melatonin levels. These 3 SNPs include CC genotype 147316, GG genotype 147379 and CC genotype 1389193 which all locate in the exon regions of ASMT and MT2 genes. Dairy goats with these SNPs have approximately 5-fold-higher melatonin levels in milk and serum than the average melatonin level detected in the current goat population. If the melatonin level impacts the milk production in goats as in cows, the results strongly suggest that these 3 SNPs can serve as the molecular markers to select the goats having the improved milk quality and yield. This is a goal of our future study.
Language	English
Publishing date	2023-03-20
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2023.1118367
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Article ; Online: Melatonin promotes the development of sheep transgenic cloned embryos by protecting donor and recipient cells.

Yao, Yujun / Yang, Ailing / Li, Guangdong / Wu, Hao / Deng, Shoulong / Yang, Hai / Ma, Wenkui / Lv, Dongying / Fu, Yao / Ji, Pengyun / Tan, Xinxing / Zhao, Wanmin / Lian, Zhengxing / Zhang, Lu / Liu, Guoshi

Cell cycle (Georgetown, Tex.)

2022 Volume 21, Issue 13, Page(s) 1360–1375

Abstract: The yield efficiency of transgenic animal generation is relatively low[1]. To improve its efficiency has become a priority task for researchers[2]. Melatonin (N-acetyl-5-methoxytryptamine, MT) is a potent-free radical scavenger and antioxidant to protect ...

Abstract	The yield efficiency of transgenic animal generation is relatively low[1]. To improve its efficiency has become a priority task for researchers[2]. Melatonin (N-acetyl-5-methoxytryptamine, MT) is a potent-free radical scavenger and antioxidant to protect mitochondria, lipids, protein and DNA from oxidative stress[3]. In this study, we observed that improving the quality of both donor and recipient cells by giving physiological concentration (10
MeSH term(s)	Animals ; Animals, Genetically Modified ; Blastocyst ; Cloning, Organism/veterinary ; Embryonic Development/physiology ; Melatonin/pharmacology ; Nuclear Transfer Techniques/veterinary ; Sheep
Chemical Substances	Melatonin (JL5DK93RCL)
Language	English
Publishing date	2022-03-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2146183-1
ISSN	1551-4005 ; 1538-4101 ; 1554-8627
ISSN (online)	1551-4005
ISSN	1538-4101 ; 1554-8627
DOI	10.1080/15384101.2022.2051122
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Article: Recent Advances of Artificial Intelligence in Cardiovascular Disease.

Chen, Zhu / Xiao, Changhu / Qiu, Haihua / Tan, Xinxing / Jin, Lian / He, Yi / Guo, Yuan / He, Nongyue

Journal of biomedical nanotechnology

2020 Volume 16, Issue 7, Page(s) 1065–1081

Abstract: Cardiovascular disease (CVD) is one of the most serious health disorders with increasing prevalence and high morbidity and mortality. Although diagnosis and treatment of CVD have achieved huge breakthrough in recent years, it still needs additional ... ...

Abstract	Cardiovascular disease (CVD) is one of the most serious health disorders with increasing prevalence and high morbidity and mortality. Although diagnosis and treatment of CVD have achieved huge breakthrough in recent years, it still needs additional enhancements, which result in the demand for new techniques. Artificial intelligence (AI) is an emerging science field that has been widely used to guide diseases diagnosis, evaluation and treatment. AI techniques are promising in CVD to explore novel pathogenic genes phenotype, guide optimal individualized therapeutic strategy, improve the management and quality of discharged patients, predict disease prognosis, and as adjuvant therapy tool. Thus, we summarize the latest application of AI techniques in clinical diagnosis, evaluation and treatment of CVD, aiming to provide novel beneficial evidence of AI and promote its application in CVD.
MeSH term(s)	Artificial Intelligence ; Cardiovascular Diseases ; Humans
Language	English
Publishing date	2020-12-11
Publishing country	United States
Document type	Journal Article
ISSN	1550-7033
ISSN	1550-7033
DOI	10.1166/jbn.2020.2955
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Article: A novel genomic approach identifies bacterial DNA-dependent RNA polymerase as the target of an antibacterial oligodeoxynucleotide, RBL1.

Tan, Xin-Xing / Chen, Yin

Biochemistry

2005 Volume 44, Issue 17, Page(s) 6708–6714

Abstract: Rapid emergence of antibiotic-resistant bacterial pathogens limits the applicability of existing drugs, which has created an urgent need for novel antibiotics preferably with entirely new mechanisms of action. Oligodeoxynucleotides (ODNs) and their ... ...

Abstract	Rapid emergence of antibiotic-resistant bacterial pathogens limits the applicability of existing drugs, which has created an urgent need for novel antibiotics preferably with entirely new mechanisms of action. Oligodeoxynucleotides (ODNs) and their modified forms have been shown to inhibit bacterial gene expression, representing a potential for developing highly specific and efficacious antibacterial agents. In this study, a tetracycline (Tet)-inducible, randomized single-stranded DNA (ssDNA) expression library was constructed and screened for conditional growth-defective or lethal phenotypes in an Escherichia coli system. From approximately 5000 transformants screened, 12 bacterial colonies were identified with either growth-defective or lethal phenotypes. One clone, CY01, with a lethal phenotype was selected and sequenced, and the ODN sequence that it generates was designated as RBL-1. Because RBL-1 shows no significant homologies to any bacterial gene sequence, a potential RBL-1 targeting protein was isolated by affinity purification. Using mass spectrometry analysis, this protein was identified as bacterial DNA-dependent RNA polymerase (RNAP). RBL-1 was further shown to effectively inhibit RNA polymerase activity in vitro. The usage of this randomized ssDNA expression library screening technology to selectively modulate production and/or function of proteins may provide a powerful strategy in both identifying novel gene targets for antibiotic discovery and developing novel antibacterial agents.
MeSH term(s)	Amino Acid Sequence ; Anti-Bacterial Agents/chemistry ; Anti-Bacterial Agents/metabolism ; DNA, Bacterial/genetics ; DNA, Bacterial/metabolism ; DNA, Single-Stranded/biosynthesis ; DNA, Single-Stranded/genetics ; DNA, Single-Stranded/metabolism ; DNA-Directed RNA Polymerases/antagonists & inhibitors ; DNA-Directed RNA Polymerases/genetics ; DNA-Directed RNA Polymerases/metabolism ; Escherichia coli/enzymology ; Escherichia coli/genetics ; Escherichia coli/growth & development ; Gene Expression Regulation, Bacterial/genetics ; Gene Library ; Gene Targeting/methods ; Genetic Vectors ; HeLa Cells ; Humans ; Molecular Sequence Data ; Moloney murine leukemia virus/genetics ; Oligodeoxyribonucleotides/chemistry ; Oligodeoxyribonucleotides/genetics ; Oligodeoxyribonucleotides/metabolism ; Protein Binding ; Tetracycline/chemistry
Chemical Substances	Anti-Bacterial Agents ; DNA, Bacterial ; DNA, Single-Stranded ; Oligodeoxyribonucleotides ; DNA-Directed RNA Polymerases (EC 2.7.7.6) ; Tetracycline (F8VB5M810T)
Language	English
Publishing date	2005-05-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	1108-3
ISSN	1520-4995 ; 0006-2960
ISSN (online)	1520-4995
ISSN	0006-2960
DOI	10.1021/bi0475626
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Article ; Online: Metatranscriptomics and pyrosequencing facilitate discovery of potential viral natural enemies of the invasive Caribbean crazy ant, Nylanderia pubens.

Valles, Steven M / Oi, David H / Yu, Fahong / Tan, Xin-Xing / Buss, Eileen A

PloS one

2012 Volume 7, Issue 2, Page(s) e31828

Abstract: Background: Nylanderia pubens (Forel) is an invasive ant species that in recent years has developed into a serious nuisance problem in the Caribbean and United States. A rapidly expanding range, explosive localized population growth, and control ... ...

Abstract	Background: Nylanderia pubens (Forel) is an invasive ant species that in recent years has developed into a serious nuisance problem in the Caribbean and United States. A rapidly expanding range, explosive localized population growth, and control difficulties have elevated this ant to pest status. Professional entomologists and the pest control industry in the United States are urgently trying to understand its biology and develop effective control methods. Currently, no known biological-based control agents are available for use in controlling N. pubens. Methodology and principal findings: Metagenomics and pyrosequencing techniques were employed to examine the transcriptome of field-collected N. pubens colonies in an effort to identify virus infections with potential to serve as control agents against this pest ant. Pyrosequencing (454-platform) of a non-normalized N. pubens expression library generated 1,306,177 raw sequence reads comprising 450 Mbp. Assembly resulted in generation of 59,017 non-redundant sequences, including 27,348 contigs and 31,669 singlets. BLAST analysis of these non-redundant sequences identified 51 of potential viral origin. Additional analyses winnowed this list of potential viruses to three that appear to replicate in N. pubens. Conclusions: Pyrosequencing the transcriptome of field-collected samples of N. pubens has identified at least three sequences that are likely of viral origin and, in which, N. pubens serves as host. In addition, the N. pubens transcriptome provides a genetic resource for the scientific community which is especially important at this early stage of developing a knowledgebase for this new pest.
MeSH term(s)	Animals ; Ants/genetics ; Ants/physiology ; Computational Biology ; DNA/analysis ; Decision Support Techniques ; Expressed Sequence Tags ; Gene Expression Profiling/methods ; Gene Library ; Metagenomics ; Microsatellite Repeats ; Pest Control, Biological ; RNA/analysis ; RNA, Messenger/metabolism ; Sequence Analysis, DNA/methods ; Transcriptome
Chemical Substances	RNA, Messenger ; RNA (63231-63-0) ; DNA (9007-49-2)
Language	English
Publishing date	2012-02-27
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0031828
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Article: Physical and genetic interactions link hox function with diverse transcription factors and cell signaling proteins.

Bondos, Sarah E / Tan, Xin-Xing / Matthews, Kathleen S

Molecular & cellular proteomics : MCP

2006 Volume 5, Issue 5, Page(s) 824–834

Abstract: Positional information provided by Hox homeotic transcription factors is integrated with other transcription factors and cell signaling cascades in specific combinations to dictate context- and gene-specific Hox activity. Protein-protein interactions ... ...

Abstract	Positional information provided by Hox homeotic transcription factors is integrated with other transcription factors and cell signaling cascades in specific combinations to dictate context- and gene-specific Hox activity. Protein-protein interactions between these groups have long been hypothesized to modulate Hox functions, yielding a context-specific function. However, difficulties in applying interaction screens to potent transcription factors have limited partner identification. A yeast two-hybrid screen using transcription activation-deficient mutants of the Drosophila melanogaster Hox protein Ultrabithorax IB identified an array of interacting proteins, consisting primarily of transcription factors and components of cell signaling pathways. Interactions were confirmed with wild-type Ultrabithorax (UBX) in phage display experiments and by immunoprecipitation for a subset of partners. In vivo assays demonstrated that two Ultrabithorax IB partners, Armadillo, regulated by Wingless/WNT signaling, and the homeodomain protein Aristaless, inhibit UBX-dependent haltere development from the default wing development pathway. Therefore, transcription factors and cell signaling proteins that subdivide Hox-specified tissues can both alter Hox function in vivo and interact with the corresponding Hox protein in vitro. UBX may also modulate partner function: the pupal death phenotype induced by ectopic expression of the UBX partner Hairy required the presence of UBX. Thus, Hox.transcription factor complexes may integrate a variety of positional cues, generating the specificity and versatility required for context-dependent Hox function.
MeSH term(s)	Amino Acid Sequence ; Animals ; Drosophila Proteins/chemistry ; Drosophila Proteins/metabolism ; Drosophila melanogaster/metabolism ; Gene Expression ; Heterozygote ; Homeodomain Proteins/chemistry ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Molecular Sequence Data ; Phenotype ; Protein Binding ; Recombinant Fusion Proteins/metabolism ; Signal Transduction ; Transcription Factors/metabolism
Chemical Substances	Drosophila Proteins ; Homeodomain Proteins ; Recombinant Fusion Proteins ; Transcription Factors
Language	English
Publishing date	2006-02-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	2075924-1
ISSN	1535-9484 ; 1535-9476
ISSN (online)	1535-9484
ISSN	1535-9476
DOI	10.1074/mcp.M500256-MCP200
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Zs.A 5599: Show issues

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Article: Peptide nucleic acid antisense oligomer as a therapeutic strategy against bacterial infection: proof of principle using mouse intraperitoneal infection.

Tan, Xin-Xing / Actor, Jeffrey K / Chen, Yin

Antimicrobial agents and chemotherapy

2005 Volume 49, Issue 8, Page(s) 3203–3207

Abstract: Antisense oligodeoxynucleotides (ODNs) and their analogs have been successfully utilized to inhibit gene expression and bacterial growth in vitro or in cell culture. In this study, acpP-targeting antisense peptide nucleic acid (PNA) and its peptide ... ...

Abstract	Antisense oligodeoxynucleotides (ODNs) and their analogs have been successfully utilized to inhibit gene expression and bacterial growth in vitro or in cell culture. In this study, acpP-targeting antisense peptide nucleic acid (PNA) and its peptide conjugate were tested as potential antibacterial agents in two groups of experiments using a mouse model. In the first group, Escherichia coli mutant strain SM101 with a defective outer membrane was used to induce bacteremia and peritonitis in BALB/c mice by intraperitoneal (i.p.) injection. The resulting bacteremia was fatal within 48 h. A single i.p injection of 5 nmol (or more) of PNA administered 30 min before bacterial challenge significantly reduced the bacterial load in mouse blood. Reductions in serum concentrations of the proinflammatory cytokines tumor necrosis factor alpha, interleukin-1beta (IL-1beta), IL-6, and IL-12 were also observed. PNA treatment was effective in rescuing 100% of infected animals. In the second group, bacteremia in BALB/c mice was induced by i.p. injection of E. coli wild-type strain K-12. The infected mice were treated by a single intravenous injection of peptide-PNA conjugate 30 min after bacterial challenge. Treatment with the peptide-PNA conjugate significantly reduced the K-12 load, with modest reduction in cytokine concentrations. The conjugate treatment was also able to rescue up to 60% of infected animals. This report is the first demonstration of ODNs' antibacterial efficacy in an animal disease model. The ability of PNA and its peptide conjugate to inhibit bacterial growth and to prevent fatal infection demonstrates the potential for this new class of antibacterial agents.
MeSH term(s)	Acyl Carrier Protein/genetics ; Acyl Carrier Protein/metabolism ; Animals ; Anti-Bacterial Agents/administration & dosage ; Anti-Bacterial Agents/metabolism ; Anti-Bacterial Agents/therapeutic use ; Apoproteins/genetics ; Apoproteins/metabolism ; Bacteremia/drug therapy ; Bacteremia/microbiology ; Bacteremia/mortality ; Dimerization ; Disease Models, Animal ; Escherichia coli/genetics ; Escherichia coli/pathogenicity ; Escherichia coli Infections/drug therapy ; Escherichia coli Infections/microbiology ; Escherichia coli Infections/mortality ; Escherichia coli K12/pathogenicity ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Fatty Acid Synthase, Type II ; Humans ; Mice ; Mice, Inbred BALB C ; Mutation ; Oligonucleotides, Antisense/administration & dosage ; Oligonucleotides, Antisense/metabolism ; Oligonucleotides, Antisense/therapeutic use ; Peptide Nucleic Acids/administration & dosage ; Peptide Nucleic Acids/metabolism ; Peptide Nucleic Acids/therapeutic use ; Peritonitis/drug therapy ; Peritonitis/microbiology ; Peritonitis/mortality ; Treatment Outcome
Chemical Substances	Acyl Carrier Protein ; Anti-Bacterial Agents ; Apoproteins ; Escherichia coli Proteins ; Oligonucleotides, Antisense ; Peptide Nucleic Acids ; acpP protein, E coli ; Fatty Acid Synthase, Type II (EC 6.-)
Language	English
Publishing date	2005-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.49.8.3203-3207.2005
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Zs.A 809: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article: Metatranscriptomics and Pyrosequencing Facilitate Discovery of Potential Viral Natural Enemies of the Invasive Caribbean Crazy Ant, Nylanderia pubens

Valles, Steven M / Boudinot, Pierre / Buss, Eileen A / Oi, David H / Tan, Xin-Xing / Yu, Fahong

PloS one. , v. 7, no. 2

2012

Abstract: Nylanderia pubens (Forel) is an invasive ant species that in recent years has developed into a serious nuisance problem in the Caribbean and United States. A rapidly expanding range, explosive localized population growth, and control difficulties have ... ...

Title variant	Metatranscriptomics of Nylanderia pubens
Abstract	Nylanderia pubens (Forel) is an invasive ant species that in recent years has developed into a serious nuisance problem in the Caribbean and United States. A rapidly expanding range, explosive localized population growth, and control difficulties have elevated this ant to pest status. Professional entomologists and the pest control industry in the United States are urgently trying to understand its biology and develop effective control methods. Currently, no known biological-based control agents are available for use in controlling N. pubens . Metagenomics and pyrosequencing techniques were employed to examine the transcriptome of field-collected N. pubens colonies in an effort to identify virus infections with potential to serve as control agents against this pest ant. Pyrosequencing (454-platform) of a non-normalized N. pubens expression library generated 1,306,177 raw sequence reads comprising 450 Mbp. Assembly resulted in generation of 59,017 non-redundant sequences, including 27,348 contigs and 31,669 singlets. BLAST analysis of these non-redundant sequences identified 51 of potential viral origin. Additional analyses winnowed this list of potential viruses to three that appear to replicate in N. pubens . Pyrosequencing the transcriptome of field-collected samples of N. pubens has identified at least three sequences that are likely of viral origin and, in which, N. pubens serves as host. In addition, the N. pubens transcriptome provides a genetic resource for the scientific community which is especially important at this early stage of developing a knowledgebase for this new pest.
MeSH term(s)	Animals ; Ants/genetics/physiology ; Computational Biology ; Decision Support Techniques ; DNA/analysis ; Expressed Sequence Tags ; Gene Expression Profiling/methods ; Gene Library ; Metagenomics ; Microsatellite Repeats ; Pest Control, Biological ; RNA/analysis ; RNA, Messenger/metabolism ; Sequence Analysis, DNA/methods ; Transcriptome
Keywords	ant control ; biological control agents ; host-pathogen relationships ; hosts ; insect colonies ; insect viruses ; invasive species ; natural enemies ; sequence analysis ; transcriptome ; viral diseases of animals and humans ; Caribbean ; United States
Language	English
Dates of publication	2012-0227
Publishing place	Public Library of Science
Document type	Article
ISSN	1932-6203
DOI	10.1371/journal.pone.0031828
Database	NAL-Catalogue (AGRICOLA)

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Article: DNA enzyme generated by a novel single-stranded DNA expression vector inhibits expression of the essential bacterial cell division gene ftsZ.

Tan, Xin-Xing / Rose, Knesha / Margolin, William / Chen, Yin

Biochemistry

2004 Volume 43, Issue 4, Page(s) 1111–1117

Abstract: Rapid emergence of antibiotic-resistant bacterial pathogens has created urgent demand for the discovery and development of new antibacterial agents directed toward novel targets. Antisense oligodeoxynucleotides (AS-ODN) and their modified forms have been ...

Abstract	Rapid emergence of antibiotic-resistant bacterial pathogens has created urgent demand for the discovery and development of new antibacterial agents directed toward novel targets. Antisense oligodeoxynucleotides (AS-ODN) and their modified forms have been utilized to block gene expression in bacterial cells, showing potential for developing highly specific and efficacious antibacterial agents. In this study, a tetracycline-regulated expression vector was developed for generating single-stranded DNA (ssDNA) of a desired target sequence in bacterial cells. This inducible ssDNA expression vector was tested for producing a DNA enzyme designed to specifically cleave ftsZ mRNA. Our results indicate that the expressed DNA enzyme molecules not only repress ftsZ gene expression and but also inhibit bacterial cell proliferation. Although we believe that the cleavage of ftsZ mRNA by the expressed DNA enzyme molecules is responsible for the inhibitory effects on ftsZ gene expression and bacterial cell proliferation, the antisense mechanism could also be responsible for the biological effects. The ability of this ssDNA expression system to selectively modulate gene expression may provide a powerful strategy in determining the contribution of a given gene product to bacterial growth or pathogenesis and opens a new venue for developing antibacterial agents.
MeSH term(s)	Bacterial Proteins/antagonists & inhibitors ; Bacterial Proteins/biosynthesis ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/antagonists & inhibitors ; Cytoskeletal Proteins/biosynthesis ; Cytoskeletal Proteins/genetics ; DNA, Catalytic/biosynthesis ; DNA, Catalytic/chemistry ; DNA, Catalytic/genetics ; DNA, Single-Stranded/biosynthesis ; DNA, Single-Stranded/chemistry ; DNA, Single-Stranded/genetics ; Down-Regulation/genetics ; Enzyme Induction/genetics ; Escherichia coli/enzymology ; Escherichia coli/genetics ; Escherichia coli/growth & development ; Gene Expression Regulation, Bacterial ; Genetic Vectors/chemical synthesis ; Growth Inhibitors/biosynthesis ; Growth Inhibitors/chemistry ; Growth Inhibitors/genetics ; Nucleic Acid Conformation ; RNA, Bacterial/chemistry ; RNA-Directed DNA Polymerase/biosynthesis ; RNA-Directed DNA Polymerase/genetics ; Tetracycline/chemistry
Chemical Substances	Bacterial Proteins ; Cytoskeletal Proteins ; DNA, Catalytic ; DNA, Single-Stranded ; FtsZ protein, Bacteria ; Growth Inhibitors ; RNA, Bacterial ; RNA-Directed DNA Polymerase (EC 2.7.7.49) ; Tetracycline (F8VB5M810T)
Language	English
Publishing date	2004-02-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	1108-3
ISSN	1520-4995 ; 0006-2960
ISSN (online)	1520-4995
ISSN	0006-2960
DOI	10.1021/bi035164h
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Article: Transcription activation by ultrabithorax Ib protein requires a predicted alpha-helical region.

Tan, Xin-Xing / Bondos, Sarah / Li, Likun / Matthews, Kathleen S

Biochemistry

2002 Volume 41, Issue 8, Page(s) 2774–2785

Abstract: Characterization of their transcription activation domains is critical to understanding functional specificity within the Hox family of proteins. However, few Hox activation domains have been identified and none characterized in detail. In this study, ... ...

Abstract	Characterization of their transcription activation domains is critical to understanding functional specificity within the Hox family of proteins. However, few Hox activation domains have been identified and none characterized in detail. In this study, promotor-reporter assays in yeast and Drosophila S2 cell culture were used to refine the boundaries of the activation domain of the Drosophila Hox protein Ultrabithorax (Ubx) and to identify critical elements within this domain. We found that residues 159-242 were sufficient for 50% function, and full transactivation capacity was achieved with inclusion of additional N-terminal sequences. Activation domain sequence and placement relative to the homeodomain differ between Ubx and other Hox proteins, consistent with the possibility that diverse activation mechanisms contribute to functional distinctions in vivo. The essential residues 159-242 in the UbxIb activation domain are predicted to contain a beta-sheet segment followed by an alpha-helix. This putative alpha-helical region was established to be necessary, but not sufficient, for transcriptional activation. Disruption of the helix by proline substitutions abolished activation function, while alteration of side chains presented on the surface of this putative helix with alanine or lysine mutations had no significant effect on activity. Collectively, these data indicate that this secondary structural element is a key component in forming an effective activation domain in the UbxIb protein. Interestingly, the alpha-helix critical for transcriptional activation is found only for Ubx orthologs from flies and not other species. The mutant Ubx proteins generated in this study have potential applications in deciphering Hox functions in vivo.
MeSH term(s)	Amino Acid Sequence ; Animals ; Cell Line ; Chloramphenicol O-Acetyltransferase/genetics ; DNA-Binding Proteins/chemistry ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; DNA-Binding Proteins/physiology ; Drosophila Proteins ; Drosophila melanogaster ; Homeodomain Proteins/chemistry ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Homeodomain Proteins/physiology ; Mutagenesis ; Protein Structure, Secondary ; Saccharomyces cerevisiae/metabolism ; Sequence Homology, Amino Acid ; Transcription Factors ; Transcriptional Activation/physiology
Chemical Substances	DNA-Binding Proteins ; Drosophila Proteins ; Homeodomain Proteins ; Transcription Factors ; Ubx protein, Drosophila ; Chloramphenicol O-Acetyltransferase (EC 2.3.1.28)
Language	English
Publishing date	2002-02-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	1108-3
ISSN	1520-4995 ; 0006-2960
ISSN (online)	1520-4995
ISSN	0006-2960
DOI	10.1021/bi011967y
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