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Generation of specific antibodies against Ki-1/57 regulatory protein for functional analysis of Ki-1/57 knockout mice

Grant number: 17/20266-6
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): March 01, 2018
Effective date (End): February 29, 2020
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Jörg Kobarg
Grantee:Carolina Colleti
Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil


Ki-1/57 protein was identified through the cross reactivity of the anti-CD30 monoclonal antibody Ki-1, in Hodgkin lymphoma cells. Studies on Ki-1/57 show that it undergoes several types of post-translational modifications (phosphorylation, methylation, SUMOylation) and that is an intrinsically unstructured protein. These and other characteristics identified, as shuttling between cytoplasm and nucleus and transcriptional regulation activity, suggests that Ki-1/57 could be an oncoprotein. It is also known that Ki-1/57 is overexpressed in several types of tumors, such as T-cell lymphoma, adenocarcinoma, prostate carcinoma and bladder carcinoma. The CGI-55 protein presents 40,7% of identity and 67,4% of similarity with Ki-1/57 and presents cytoplasmic and nuclear localization as well, indicating that both may be paralogs proteins and, in addition, may have similar/redundant functions in the cells. Yeast two-hybrid and Affymetrix microarray experiments performed by the group, identified that the expression of 413 genes was altered by Ki-1/57 overexpression (with 88% of them down-regulated). The most part of these altered genes are related to cell cycle, proliferation and apoptosis, which indicates their involvement in mechanisms of cellular response to stress. These data together with available information about protein-protein interaction and subcellular location, suggests that Ki-1/57 activity, under these conditions, would occur at different levels of gene expression control, from transcription to translation. Based on the Ki-1/57 involvement in proliferation and cell death pathways and because of the possibility of being an oncoprotein, an in-depth study of their cellular role is needed. Prof. Jörg Kobarg Lab produced knockout mice for the Ki-1/57 gene by CRISPR/Cas9 technique and, as scope of this project, it is proposed to comprehend the role of Ki-1/57 in the cell. This will be done through comparative analysis of tissues of knockout mice compared to the wild, as well as carry out cell studies with immortalized cells of the knockout and wild mice. (AU)

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
COLLETI, Carolina. HABP4 and colon cancer: characterization of the HABP4 role in tumor cells and animal model. 2020. Master's Dissertation - Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Farmacêuticas Campinas, SP.

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