Quantify the basic needs of cell division
Understanding a biological process requires analyzing the genes and proteins involved. However, quantifying a certain protein component in a critical structure is not easy. Fortunately, scientists at the IGC (Instituto Gulbenkian de Ciência) in Portugal have solved this problem. They studied the centromere of human cells by fluorescence technology and found that approximately 400 CENP-A molecules were required for centromere formation.
Centromere is an important protein structure on the chromosome, which can recruit related molecular machinery to distribute chromosomes to two daughter cells, and this process is the basis of cell division. If the orientation of the centromere changes, or if the protein that makes up the centromere is destroyed, cell division will be abnormal. It has been known that the protein CENP-A is extremely important for the function of the centromere, but it is unclear how many CENP-A molecules are present in the centromere. Such information will help people understand the establishment and maintenance mechanisms of centromeres.
"We know that CENP-A plays a crucial role in the centromere formation process. Previous studies have shown that the lack of this protein cell can not be correctly split, the number of chromosomes passed to the daughter cells will be affected. How much CENP-A does silk formation need? We need to find ways to quantify this nanoscale molecule," explains Dani Bodor, the first author of the article.
The research team used genetic engineering technology to fuse a gene encoding a fluorescent protein with the CENP-A gene to fluoresce all CENP-A proteins produced by the cell. They then observed the cells under a microscope and quantified the fluorescence at the entire cell and centromere. They found that about 400 CENP-A molecules in human cells appear at the centromere. The researchers pointed out that the above methods can also be used to solve other biological problems.
“Similar technology was originally used for yeast. No one has used it for more complicated cells before, but we have gained some inspiration. The shape and size of yeast cells are similar, but the morphology and size of human cells are quite different. , increasing the complexity of using such technologies," said Dani Bodor.
Finally, the researchers used two other techniques to validate the results. Studies have shown that no matter which method is used, there are about 400 CENP-A molecules at the centromere.
"The centromere requires a very stable structure to ensure that the chromosomes are correctly delivered to the daughter cells during cell division. At the time of cell division, CENP-A protein is also transmitted to the daughter cells, but the number of molecules received by each cell is not The same. With 400 molecules in the cell, you can ensure that enough CENP-A is passed down to form centromeres. Determining the number of CENP-A helps us understand the formation and inheritance of centromeres."
“Today, more and more laboratories are beginning to address quantitative problems in biology. And our approach is very useful in this regard,†Lars said.
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