mCSPP may play an important role in cell cycle progression and cell division. The identification of CSPP in mice and the study of its function was first published in 2009. Scientists successfully cloned the cDNA for two alternatively spliced forms of mCSPP: mCSPP-1, and mCSPP-2. They determined their cellular localization with GFP-tagged mCSPP-1, and -2 (see Fig.1). The image shows that GFP-mCSPP-1 localized to the spindle pole and central spindle. GFP-mCSPP-2 also localized to the central spindle in anaphase. However, GFP-CSPP-2 predominantly localized to the spindle microtubules in metaphase. Moreover, it was reported that mCSPP interact with MyoGEF, a guanine nucleotide exchange factor that localizes to the central spindle and cleavage furrow. Depletion of mCSPP or MyoGEF by RNA-interference will causes defects in mitosis and cytokinesis, such as metaphase arrest and furrow regression. We know that mCSPP is alternatively spliced and some alternatively spliced variants have different functions in cell cycle progression and cell division.
Figure 1-The localization of mCSPP-1.
Figure 2-The structure and alternative splicing pattern of mCSPP gene.
Figure 3-The position of each primer pairs and the size of products.
Figure 4-RT-PCR and sequencing results with RNA from mouse kidney (lane 1-6), lung (lane 7-12), and heart (lane13-18). Six primer pairs(including mGAPDH) were used.
Centrosome/Spindle Pole-associated Protein (CSPP) was first identified by Dr. Sebastian Patzke and Dr. Trond Stokke in 2006. It is a protein that is concentrated at the spindle pole and central spindle during mitosis and cytokinesis and plays an important role in cell cycle progression. In 2009, scientists discovered the expression of CSPP in mouse and identified several different isoforms of mouse CSPP (mCSPP) such as mCSPP-1 and mCSPP-2. These isoforms are the result of alternative splicing of mCSPP. As of now, there is no report of the detection of tissue specific alternative splicing of mouse CSPP. In this study, we detected the alternative splicing of exon 18 in the mouse CSPP using RT-PCR. In this study, we found transcripts of mCSPP that both contained and lacked exon 18 in the kidney and lung tissue but only transcripts that contain exon 18 in heart tissue.
I would like to thank Dr. Wei for providing many kinds of mouse tissues and mRNA samples as well as useful suggestions. I would like to thank Anthony Evans and Farria Fasih-Ahmad for their fantastic work as Teaching Assistants and for their never-ending patience and efforts they put in to make this project undergo successfully. Finally, I would like to thank Dr. Rubin for his guidance and for making this project possible.
|This document was last modified 05/15/2018.|
This site is powered by the versatile Zope platform.
|This is a project of the Biology Department of Fordham University