NDLI: Structural Prediction and Mutational Analysis of the Gifsy-1Xis Protein

Content Provider	Springer Nature : BioMed Central
Author	Flanigan, Asa Gardner, Jeffrey
Abstract	Background The Gifsy-1 phage integrates into the Salmonella Typhimurium chromosome via an integrase mediated, site-specific recombination mechanism. Excision of the Gifsy-1 phage requires three proteins, the Gifsy-1 integrase (Int), the Gifsy-1 excisionase (Xis) protein, and host encoded Integration Host Factor (IHF). The Gifsy-1 xis gene encodes the 94-residue Gifsy-1 excisionase protein that has a molecular weight of 11.2 kDa and a pI of 10.2. Electrophoretic Mobility Shift Assays (EMSA) suggested at least one region of the protein is responsible for protein-DNA interactions with a tripartite DNA binding site composed of three direct imperfect repeats. Results Here we have undertaken experiments to dissect and model the structural motifs of Gifsy-1 Xis necessary for its observed DNA binding activity. Diethyl sulfate mutagenesis (DES) and mutagenic PCR techniques were used to generate Gifsy-1 xis mutants. Mutant Xis proteins that lacked activity in vivo were purified and tested by EMSA for binding to the Gifsy-1 Xis attP attachment site. Results from mutagenesis experiments and EMSA were compared to results of structural predictions and sequence analyses. Conclusion Sequence comparisons revealed evidence for three distinct structural motifs in the Gifsy-1 Xis protein. Multiple sequence alignments revealed unexpected homologies between the Gifsy-1 Xis protein and two distinct subsets of polynucleotide binding proteins. Our data may suggest a role for the Gifsy-1 Xis in the regulation of the Gifsy-1 phage excision beyond that of DNA binding and possible interactions with the Gifsy-1 Int protein.
Related Links	https://bmcmicrobiol.biomedcentral.com/counter/pdf/10.1186/1471-2180-8-199.pdf
Ending Page	12
Page Count	12
Starting Page	1
File Format	HTM / HTML
ISSN	14712180
DOI	10.1186/1471-2180-8-199
Journal	BMC Microbiology
Issue Number	1
Volume Number	8
Language	English
Publisher	BioMed Central
Publisher Date	2008-11-17
Access Restriction	Open
Subject Keyword	Microbiology Biological Microscopy Mycology Parasitology Virology Life Sciences Electrophoretic Mobility Shift Assay Luria Broth Limited Proteolysis Integration Host Factor Helix Turn Helix
Content Type	Text
Resource Type	Article
Subject	Microbiology Microbiology (medical)
Journal Impact Factor	4/2023
5-Year Journal Impact Factor	4.6/2023

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Isolation and characterization of Ehrlichia chaffeensis RNA polymerase and its use in evaluating p28 outer membrane protein gene promoters

DNA-binding by Haemophilus influenzae and Escherichia coli YbaB, members of a widely-distributed bacterial protein family

Comparative transcriptomics in Yersinia pestis: a global view of environmental modulation of gene expression

Characterization of Zur-dependent genes and direct Zur targets in Yersinia pestis

Identification and characterization of PhbF: A DNA binding protein with regulatory role in the PHB metabolism of Herbaspirillum seropedicae SmR1

Molecular characterization of Legionella pneumophila-induced interleukin-8 expression in T cells

NF-κB activation by Helicobacter pylorirequires Akt-mediated phosphorylation of p65

Staphylococcus aureus immunodominant surface antigen B is a cell-surface associated nucleic acid binding protein

RsmW, Pseudomonas aeruginosa small non-coding RsmA-binding RNA upregulated in biofilm versus planktonic growth conditions

Structural Prediction and Mutational Analysis of the Gifsy-1Xis Protein

Similar Documents

Isolation and characterization of Ehrlichia chaffeensis RNA polymerase and its use in evaluating p28 outer membrane protein gene promoters

DNA-binding by Haemophilus influenzae and Escherichia coli YbaB, members of a widely-distributed bacterial protein family

Comparative transcriptomics in Yersinia pestis: a global view of environmental modulation of gene expression

Characterization of Zur-dependent genes and direct Zur targets in Yersinia pestis

Identification and characterization of PhbF: A DNA binding protein with regulatory role in the PHB metabolism of Herbaspirillum seropedicae SmR1

Molecular characterization of Legionella pneumophila-induced interleukin-8 expression in T cells

NF-κB activation by Helicobacter pylorirequires Akt-mediated phosphorylation of p65

Staphylococcus aureus immunodominant surface antigen B is a cell-surface associated nucleic acid binding protein

RsmW, Pseudomonas aeruginosa small non-coding RsmA-binding RNA upregulated in biofilm versus planktonic growth conditions

Structural Prediction and Mutational Analysis of the Gifsy-1Xis Protein