A Mutant Stat5b with Weaker DNA Binding Affinity Defines a Key Defective Pathway in Nonobese Diabetic Mice

Abdoreza Davoodi-Semiromi, Malini Laloraya, G. Pradeep Kumar, Sharad B Purohit, Rajesh Kumar Jha, Jin-Xiong She

Research output: Contribution to journalArticle

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Abstract

A number of cytokines that finely regulate immune response have been implicated in the pathogenesis or protection of type 1 diabetes and other autoimmune diseases. It is, therefore, of pivotal importance to examine a family of proteins that serve as signal transducers and activators of transcription (STATs), which regulate the transcription of a variety of cytokines. We report here a defective gene (Stat5b) located on chromosome 11 within a previously mapped T1D susceptibility interval (Idd4) in the nonobese diabetic (NOD) mice. Our sequencing analysis revealed a unique mutation C1462A that results in a leucine to methionine (L327M) in Stat5b of NOD mice. Leu 327 , the first residue in the DNA binding domain of STAT proteins, is conserved in all identified mammalian STAT proteins. Homology modeling predicted that the mutant Stat5b has a weaker DNA binding, which was confirmed by DNA-protein binding assays. The inapt transcriptional regulation ability of the mutated Stat5b is proved by decreased levels of RNA of Stat5b-regulated genes (IL-2Rβ and Pim1). Consequently, IL-2Rβ and Pim1 proteins were shown by Western blotting to have lower levels in NOD compared with normal B6 mice. These proteins have been implicated in immune regulation, apoptosis, activation-induced cell death, and control of autoimmunity. Therefore, the Stat5b pathway is a key molecular defect in NOD mice.

Original languageEnglish (US)
Pages (from-to)11553-11561
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number12
DOIs
StatePublished - Mar 19 2004

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Inbred NOD Mouse
STAT Transcription Factors
Transcription
DNA
Genes
Cytokines
Chromosomes, Human, Pair 11
Proteins
DNA-Binding Proteins
Cell death
Chromosomes
Medical problems
Transducers
Autoimmunity
Type 1 Diabetes Mellitus
Leucine
Methionine
Autoimmune Diseases
Assays
Cell Death

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A Mutant Stat5b with Weaker DNA Binding Affinity Defines a Key Defective Pathway in Nonobese Diabetic Mice. / Davoodi-Semiromi, Abdoreza; Laloraya, Malini; Kumar, G. Pradeep; Purohit, Sharad B; Jha, Rajesh Kumar; She, Jin-Xiong.

In: Journal of Biological Chemistry, Vol. 279, No. 12, 19.03.2004, p. 11553-11561.

Research output: Contribution to journalArticle

Davoodi-Semiromi, Abdoreza ; Laloraya, Malini ; Kumar, G. Pradeep ; Purohit, Sharad B ; Jha, Rajesh Kumar ; She, Jin-Xiong. / A Mutant Stat5b with Weaker DNA Binding Affinity Defines a Key Defective Pathway in Nonobese Diabetic Mice. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 12. pp. 11553-11561.
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