{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/86725"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/86725","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Integration Host Factor and Its Interaction With the H-Prime Site of Bacteriophage Lambda","abstract":"Integration Host Factor (IHF) is a sequence-specific DNA-bending protein of Escherichia coli that recognizes the DNA through contacts made to the backbone and the minor groove. IHF binds specifically to the H$\\sp\\prime$ site, $5\\sp\\prime$-TAAAAAAGCATTGCTTATCAATTTGTTGCAACGA-$3\\sp\\prime$, with a $\\rm K\\sb{d}(app)$ of 1 nM. Substitution of an A for the G within the TTG element, which is allowable in the consensus, increases the $\\rm K\\sb{d}(app)$ to 10 nM and indicates that context is important in the recognition of this conserved position. Any other single base substitution within the TTG element increases the Kd(app) of IHF for the H$\\sp\\prime$ site to ${>}2750$ nM and shows that the interaction of IHF with the TTG sequence is very specific. By combining the genetic data of these studies with the structural data of the IHF-H$\\sp\\prime$ site cocrystal the specificity can be explained as a combination of direct and indirect readout of sequence information. Mutants that remove the glutamic acid 44 of the $\\beta$-subunit restore wild-type levels of IHF binding to an H$\\sp\\prime$ site containing a TAG element. These mutants still bind the wild-type TTG-containing H$\\sp\\prime$ site efficiently. However, these mutants do not allow IHF to recognize other mutant variants of the H$\\sp\\prime$ site. Substitution with a variety of amino acid sidechains demonstrates that both oxygens of the glutamic acid sidechain are required for full discrimination against the TAG variant. In the cocrystal, the oxygens of the glutamic acid make bonds with two flanking arginines but do not contact the DNA. These results also demonstrate that IHF can distinguish between a T:A and A:T basepair through interactions in the minor groove.","abstract_html":"Integration Host Factor (IHF) is a sequence-specific DNA-bending protein of Escherichia coli that recognizes the DNA through contacts made to the backbone and the minor groove. IHF binds specifically to the H$\\sp\\prime$ site, $5\\sp\\prime$-TAAAAAAGCATTGCTTATCAATTTGTTGCAACGA-$3\\sp\\prime$, with a $\\rm K\\sb{d}(app)$ of 1 nM. Substitution of an A for the G within the TTG element, which is allowable in the consensus, increases the $\\rm K\\sb{d}(app)$ to 10 nM and indicates that context is important in the recognition of this conserved position. Any other single base substitution within the TTG element increases the Kd(app) of IHF for the H$\\sp\\prime$ site to ${&gt;}2750$ nM and shows that the interaction of IHF with the TTG sequence is very specific. By combining the genetic data of these studies with the structural data of the IHF-H$\\sp\\prime$ site cocrystal the specificity can be explained as a combination of direct and indirect readout of sequence information. Mutants that remove the glutamic acid 44 of the <span class=\"etd-inline-math\">&beta;</span>-subunit restore wild-type levels of IHF binding to an H$\\sp\\prime$ site containing a TAG element. These mutants still bind the wild-type TTG-containing H$\\sp\\prime$ site efficiently. However, these mutants do not allow IHF to recognize other mutant variants of the H$\\sp\\prime$ site. Substitution with a variety of amino acid sidechains demonstrates that both oxygens of the glutamic acid sidechain are required for full discrimination against the TAG variant. In the cocrystal, the oxygens of the glutamic acid make bonds with two flanking arginines but do not contact the DNA. These results also demonstrate that IHF can distinguish between a T:A and A:T basepair through interactions in the minor groove.","abstract_has_math":true,"creators":["Read, Erik Karl"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":["Gardner, Jeffrey F."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-09-28T15:17:38Z","date_published":"2015-09-28T15:17:38Z","updated_at":"2026-07-22T22:26:27Z","subjects":["Biology, Molecular"],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(MiAaPQ)AAI9812746"],"render_values":[{"text":"(MiAaPQ)AAI9812746","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/86725","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gardner, Jeffrey F."]},{"key":"dc:creator","label":"Author","values":["Read, Erik Karl"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2015-09-28T15:17:38Z","10000-01-01","1997"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Molecular"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/86725","(MiAaPQ)AAI9812746"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Integration Host Factor (IHF) is a sequence-specific DNA-bending protein of Escherichia coli that recognizes the DNA through contacts made to the backbone and the minor groove. IHF binds specifically to the H$\\sp\\prime$ site, $5\\sp\\prime$-TAAAAAAGCATTGCTTATCAATTTGTTGCAACGA-$3\\sp\\prime$, with a $\\rm K\\sb{d}(app)$ of 1 nM. Substitution of an A for the G within the TTG element, which is allowable in the consensus, increases the $\\rm K\\sb{d}(app)$ to 10 nM and indicates that context is important in the recognition of this conserved position. Any other single base substitution within the TTG element increases the Kd(app) of IHF for the H$\\sp\\prime$ site to ${>}2750$ nM and shows that the interaction of IHF with the TTG sequence is very specific. By combining the genetic data of these studies with the structural data of the IHF-H$\\sp\\prime$ site cocrystal the specificity can be explained as a combination of direct and indirect readout of sequence information. Mutants that remove the glutamic acid 44 of the $\\beta$-subunit restore wild-type levels of IHF binding to an H$\\sp\\prime$ site containing a TAG element. These mutants still bind the wild-type TTG-containing H$\\sp\\prime$ site efficiently. However, these mutants do not allow IHF to recognize other mutant variants of the H$\\sp\\prime$ site. Substitution with a variety of amino acid sidechains demonstrates that both oxygens of the glutamic acid sidechain are required for full discrimination against the TAG variant. In the cocrystal, the oxygens of the glutamic acid make bonds with two flanking arginines but do not contact the DNA. These results also demonstrate that IHF can distinguish between a T:A and A:T basepair through interactions in the minor groove.","Made available in DSpace on 2015-09-28T15:17:38Z (GMT). No. of bitstreams: 2 license.txt: 4848 bytes, checksum: 96035ab3f5e1c23cc7138a224ce498bd (MD5) 9812746.pdf: 4444202 bytes, checksum: c918f5f09adfb13f3e85fdb54fbaa4b5 (MD5) Previous issue date: 1997","Embargo set by: Seth Robbins for item 88006 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","99 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1997."]},{"key":"dc:title","label":"Title","values":["Integration Host Factor and Its Interaction With the H-Prime Site of Bacteriophage Lambda"]}]}],"canonical_facts":{"dc:contributor":["Gardner, Jeffrey F."],"dc:creator":["Read, Erik Karl"],"dc:date":["2015-09-28T15:17:38Z","10000-01-01","1997"],"dc:description":["Integration Host Factor (IHF) is a sequence-specific DNA-bending protein of Escherichia coli that recognizes the DNA through contacts made to the backbone and the minor groove. IHF binds specifically to the H$\\sp\\prime$ site, $5\\sp\\prime$-TAAAAAAGCATTGCTTATCAATTTGTTGCAACGA-$3\\sp\\prime$, with a $\\rm K\\sb{d}(app)$ of 1 nM. Substitution of an A for the G within the TTG element, which is allowable in the consensus, increases the $\\rm K\\sb{d}(app)$ to 10 nM and indicates that context is important in the recognition of this conserved position. Any other single base substitution within the TTG element increases the Kd(app) of IHF for the H$\\sp\\prime$ site to ${>}2750$ nM and shows that the interaction of IHF with the TTG sequence is very specific. By combining the genetic data of these studies with the structural data of the IHF-H$\\sp\\prime$ site cocrystal the specificity can be explained as a combination of direct and indirect readout of sequence information. Mutants that remove the glutamic acid 44 of the $\\beta$-subunit restore wild-type levels of IHF binding to an H$\\sp\\prime$ site containing a TAG element. These mutants still bind the wild-type TTG-containing H$\\sp\\prime$ site efficiently. However, these mutants do not allow IHF to recognize other mutant variants of the H$\\sp\\prime$ site. Substitution with a variety of amino acid sidechains demonstrates that both oxygens of the glutamic acid sidechain are required for full discrimination against the TAG variant. In the cocrystal, the oxygens of the glutamic acid make bonds with two flanking arginines but do not contact the DNA. These results also demonstrate that IHF can distinguish between a T:A and A:T basepair through interactions in the minor groove.","Made available in DSpace on 2015-09-28T15:17:38Z (GMT). No. of bitstreams: 2 license.txt: 4848 bytes, checksum: 96035ab3f5e1c23cc7138a224ce498bd (MD5) 9812746.pdf: 4444202 bytes, checksum: c918f5f09adfb13f3e85fdb54fbaa4b5 (MD5) Previous issue date: 1997","Embargo set by: Seth Robbins for item 88006 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","99 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1997."],"dc:identifier":["http://hdl.handle.net/2142/86725","(MiAaPQ)AAI9812746"],"dc:language":["eng"],"dc:subject":["Biology, Molecular"],"dc:title":["Integration Host Factor and Its Interaction With the H-Prime Site of Bacteriophage Lambda"],"dc:type":["text"],"thesis:degree_discipline":["Microbiology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:27Z"}