Understanding biological processes often involves studying the genes and proteins that drive them. However, measuring the exact number of specific proteins in critical cellular structures can be quite challenging. Fortunately, a team of scientists at the Instituto Gulbenkian de Ciência (IGC) in Portugal has made a breakthrough. By using advanced fluorescence technology, they were able to determine that approximately 400 CENP-A molecules are needed for centromere formation in human cells.
The centromere is a vital structure on chromosomes that helps guide the accurate distribution of genetic material during cell division. It acts as a platform for molecular machinery that ensures each daughter cell receives the correct number of chromosomes. If the centromere's orientation changes or if its key proteins are disrupted, the entire process can go wrong, leading to serious consequences like chromosomal abnormalities. While it’s long been known that CENP-A plays a crucial role in centromere function, the exact number of these molecules has remained unclear—until now.
“We know that CENP-A is essential for centromere formation,†said Dani Bodor, the study’s lead author. “Without it, cells can’t divide properly, and the number of chromosomes passed to daughter cells becomes unpredictable. But how many CENP-A molecules are actually needed? We had to find a way to measure this at the nanoscale.â€
To answer this question, the researchers used genetic engineering to fuse a fluorescent protein with the CENP-A gene. This allowed them to track all the CENP-A proteins produced by the cell under a microscope. By analyzing the fluorescence intensity across the entire cell and specifically at the centromeres, they found that around 400 CENP-A molecules are present in human cells at the centromere. The technique they developed could also be applied to other biological questions, offering new ways to quantify proteins in complex systems.
“This method was previously used in yeast, but no one had applied it to more complex organisms like humans before,†explained Bodor. “Yeast cells are simpler in size and shape, while human cells are much more variable, which made this task more difficult.â€
To confirm their findings, the team used two additional techniques, and in every case, the number of CENP-A molecules at the centromere came out to about 400. “The centromere needs a stable structure to ensure that chromosomes are correctly distributed during cell division,†said Bodor. “CENP-A is passed down to daughter cells, but not all cells receive the same amount. Having 400 molecules ensures that enough is inherited to maintain functional centromeres.â€
“As more labs focus on quantitative biology, our approach offers a powerful tool for understanding how molecular components work together in living systems,†added Lars, another researcher involved in the study. “This discovery brings us one step closer to unraveling the mysteries of chromosome inheritance and cell division.â€
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