Interaction of PRP4 with Kruppel-like factor 13 regulates CCL5 transcription.
Activation of resting T lymphocytes initiates differentiation into mature effector cells over 3-7 days. The chemokine CCL5 (RANTES) and its major transcriptional regulator, Krüppel-like factor 13 (KLF13), are expressed late (3-5 days) after activation in T lymphocytes. Using yeast two-hybrid screening of a human thymus cDNA library, PRP4, a serine/threonine protein kinase, was identified as a KLF13-binding protein. Specific interaction of KLF13 and PRP4 was confirmed by reciprocal coimmunoprecipitation. PRP4 is expressed in PHA-stimulated human T lymphocytes from days 1 and 7 with a peak at day 3. Using an in vitro kinase assay, it was found that PRP4 phosphorylates KLF13. Furthermore, although phosphorylation of KLF13 by PRP4 results in lower binding affinity to the A/B site of the CCL5 promoter, coexpression of PRP4 and KLF13 increases nuclear localization of KLF13 and CCL5 transcription. Finally, knock-down of PRP4 by small interfering RNA markedly decreases CCL5 expression in T lymphocytes. Thus, PRP4-mediated phosphorylation of KLF13 plays a role in the regulation of CCL5 expression in T lymphocytes.
Huang
Boli
B
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Ahn
Yong-Tae
YT
McPherson
Lisa
L
Clayberger
Carol
C
Krensky
Alan M
AM
D021581:Active Transport, Cell Nucleus / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D019742:Chemokines, CC / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation / Q000276:immunology*; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D016176:T-Lymphocyte Subsets / Q000201:enzymology / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
C483024:CCL5 protein, human; D018797:Cell Cycle Proteins; D018946:Chemokine CCL5; D019742:Chemokines, CC; C486423:KLF13 protein, human; D051741:Kruppel-Like Transcription Factors; D012097:Repressor Proteins; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; C489984:PRPF4B protein, human; D017346:Protein Serine-Threonine Kinases; D020928:Mitogen-Activated Protein Kinases