General notesab94115 is a 293T cell transfected lysate in which Human Bub1 has been transiently over-expressed using a pCMV-Bub1 plasmid. The lysate is provided in 1X Sample Buffer.
Note: For more details about how the transfected lysate was prepared view preparation notes
BackgroundFunction: Serine/threonine-protein kinase that performs 2 crucial functions during mitosis: it is essential for spindle-assembly checkpoint signaling and for correct chromosome alignment. Has a key role in the assembly of checkpoint proteins at the kinetochore, being required for the subsequent localization of CENPF, BUB1B, CENPE and MAD2L1. Required for the kinetochore localization of PLK1. Plays an important role in defining SGOL1 localization and thereby affects sister chromatid cohesion. Acts as a substrate for anaphase-promoting complex or cyclosome (APC/C) in complex with its activator CDH1 (APC/C-Cdh1). Necessary for ensuring proper chromosome segregation and binding to BUB3 is essential for this function. Can regulate chromosome segregation in a kinetochore-independent manner. Can phosphorylate BUB3. The BUB1-BUB3 complex plays a role in the inhibition of APC/C when spindle-assembly checkpoint is activated and inhibits the ubiquitin ligase activity of APC/C by phosphorylating its activator CDC20. This complex can also phosphorylate MAD1L1. Kinase activity is essential for inhibition of APC/CCDC20 and for chromosome alignment but does not play a major role in the spindle-assembly checkpoint activity. Mediates cell death in response to chromosome missegregation and acts to suppress spontaneous tumorigenesis.
Tissue specificity: High expression in testis and thymus, less in colon, spleen, lung and small intestine. Expressed in fetal thymus, bone marrow, heart, liver, spleen and thymus. Expression is associated with cells/tissues with a high mitotic index.
Similarity: Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain.
Domain: The KEN box is required for its ubiquitination and degradation.
BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
PTM: Phosphorylated upon DNA damage, probably by ATM or ATR. Upon spindle-assembly checkpoint activation it is hyperphosphorylated and its kinase activity toward CDC20 is stimulated. Phosphorylation at Thr-609 is required for interaction with PLK1, phosphorylation at this site probably creates a binding site for the POLO-box domain of PLK1, thus enhancing the PLK1-BUB1 interaction.
Ubiquitinated and degraded during mitotic exit by APC/C-Cdh1.