Identification of non-histone protein targets of KMT5A in hematopoietic cells.

Teniola Idowu

Document Type

Article

Publication Date

Summer 2017

JAX Location

In: Student Reports, Summer 2017, Jackson Laboratory

Abstract

The regulation of hematopoietic cell differentiation and proliferation is critical for the proper development of the cellular components of blood. The loss of this hematopoietic regulation has been implicated in several blood cancers, neoplasia, and neurodegenerative diseases. 1 ,2 Many epigenetic factors aid in this regulation and a particular enzyme, KMT5A, has been shown to play a role in the differentiation and proliferation of cells during hematopoiesis. Recent work in the Trowbridge lab has shown that Kmt5a knockdown cause the expansion of myeloid cells during differentiation, but the mechanism behind this is still undetermined. This project showed that the knockdown of Kmt5a causes alterations in the transcriptional activity of the KMT5A non-histone protein targets NUMB, P53 and PCNA. A lentiviral knockdown approach was used to reduce Kmt5a expression in MPP4 cells, which were then transplanted in B6 mice. After a 2 and 4 week period the mice were peripherally bled, and the cells were sorted using flow cytometry. RNA extraction and cDNA synthesis was performed using MPP4 cells. The cDNA was used in RT-PCR with primers for the downstream targets of NUMB, P53 and PCNA to show that KMT5A methylation plays a role in the expansion of myeloid cells. Ultimately, the insight this project provides on the mechanism underlying myeloid expansion can lead to the creation of more specific targets for cancer therapies that do not damage normal hematopoietic cells.

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