Cloning, overexpression and characterization of iron regulatory proteins from insects

Persistent Link:
http://hdl.handle.net/10150/279922
Title:
Cloning, overexpression and characterization of iron regulatory proteins from insects
Author:
Zhang, Dianzheng
Issue Date:
2001
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Iron is essential for life and iron homeostasis is important for all species. Compared to the understanding of iron metabolisms in vertebrates, we know much less about insect intracellular iron homeostasis. The iron regulatory proteins (IRPs) play central roles in this process by interaction with iron responsive elements (IREs). Here, I report the cloning, sequencing, overexpression, purification and characterization of IRP1s from two insect species, Manduca sexta and Aedes aegypti. Electrophoretic mobility shift assays demonstrated that both IRP1s specifically bind IREs s not only from the same species, but also from human ferritin IRE. Another ferritin subunit also was cloned from Manduca sexta and an IRE was identified in the 5'-untranslated region of the mRNA, and the IRE reacted with Manduca IRP1 specifically. Transcription/translation assays demonstrated that both IRP1s repress ferritin synthesis in vitro, and the repression is IRE dependent. Iron administration to Manduca sexta increased hemolymph ferritin levels and decreased fat body IRP1/IRE binding activities without affecting either the IRP1 mRNA or protein levels. These data indicates that translational control of ferritin synthesis by IRP1/IRE interaction could occur in insects in a manner similar to that of mammals. To our knowledge this is the first report of the control of insect ferritin synthesis by IRP1/IRE interaction. The different responses to reducing agent of Manduca sexta and mammalian IRP1s could provide a potential future strategy for designing pesticides in insect control.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Biology, Molecular.; Biology, Entomology.; Biology, Animal Physiology.
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Graduate College; Nutritional Sciences
Degree Grantor:
University of Arizona
Advisor:
Winzerling, Joy J.

Full metadata record

DC FieldValue Language
dc.language.isoen_USen_US
dc.titleCloning, overexpression and characterization of iron regulatory proteins from insectsen_US
dc.creatorZhang, Dianzhengen_US
dc.contributor.authorZhang, Dianzhengen_US
dc.date.issued2001en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractIron is essential for life and iron homeostasis is important for all species. Compared to the understanding of iron metabolisms in vertebrates, we know much less about insect intracellular iron homeostasis. The iron regulatory proteins (IRPs) play central roles in this process by interaction with iron responsive elements (IREs). Here, I report the cloning, sequencing, overexpression, purification and characterization of IRP1s from two insect species, Manduca sexta and Aedes aegypti. Electrophoretic mobility shift assays demonstrated that both IRP1s specifically bind IREs s not only from the same species, but also from human ferritin IRE. Another ferritin subunit also was cloned from Manduca sexta and an IRE was identified in the 5'-untranslated region of the mRNA, and the IRE reacted with Manduca IRP1 specifically. Transcription/translation assays demonstrated that both IRP1s repress ferritin synthesis in vitro, and the repression is IRE dependent. Iron administration to Manduca sexta increased hemolymph ferritin levels and decreased fat body IRP1/IRE binding activities without affecting either the IRP1 mRNA or protein levels. These data indicates that translational control of ferritin synthesis by IRP1/IRE interaction could occur in insects in a manner similar to that of mammals. To our knowledge this is the first report of the control of insect ferritin synthesis by IRP1/IRE interaction. The different responses to reducing agent of Manduca sexta and mammalian IRP1s could provide a potential future strategy for designing pesticides in insect control.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectBiology, Molecular.en_US
dc.subjectBiology, Entomology.en_US
dc.subjectBiology, Animal Physiology.en_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineNutritional Sciencesen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorWinzerling, Joy J.en_US
dc.identifier.proquest3040137en_US
dc.identifier.bibrecord.b4248196xen_US
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