Playing metabolism tag

Welcome to …(drumroll here)…  Acetyl Tag, the new game that goes beyond the cellular nucleus to the cytoplasm, to a place where few would guess finding — let alone imagine — masses of acetylated metabolic enzymes.     

“We have discovered an entirely new layer of control of metabolism,” says Yue Xiong, Ph.D., professor of biochemistry and biophysics and a member of the UNC Lineberger Comprehensive Cancer Center.

Xiong points out that almost all previous studies on acetylation have focused on proteins in the nucleus, where acetyl tags on proteins regulate how tightly the DNA’s genetic code is packaged.  It’s known that protein acetylation plays a key role in gene expression.

But  Xiong along with study co-leader Kun-Liang Guan, professor of pharmacology at the University of California, San Diego, wanted to determine if acetylation also plays a role in the other half of the cell, the cytoplasm.

After separating the nucleus and the cytoplasm of human primary liver cells, the study team used mass spectroscopy to take a chemical census of the cytoplasm’s contents .

And to their surprise, they identified approximately a thousand new proteins that are acetylated, greatly expanding the previously recognized repertoire of fewer than one hundred.  Nearly every metabolic enzyme was acetylated, presumably because their starting material was liver, an organ rich in metabolic activity.

In addition, the researchers discovered that blocking acetylation chemically or genetically affected these metabolic enzymes in a number of different ways, either by stimulating its activity, inhibiting it, or degrading the protein itself. They suspect that acetylation is important for coordinating not only the players within a metabolic pathway but also between different pathways.

 The next step is to take their finding in normal cells and see how it can inform their study of tumor cells. The researchers are in the process of looking at each metabolic enzyme, one-by-one, to see which one displays the most disparate acetylation patterns between normal and cancer cells. They will then try to use the very same proteins that tack on or pull off those acetyl groups – called acetylases or deacetylases, respectively — to modify acetylation and thwart cancer development.

“If we can identify which enzyme or enzymes are responsible for the difference in metabolism between normal and tumor cells, then we may have  have new targets for the treating cancer patients,” says Xiong.

The study appears in the February 19 issue of  Science.

Les Lang

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Islet Package Hot spots

Despite the wintry chill down here in the land of hope and hominy,  the title of this blog post has niente to do with a travel offer  to warmer weather.

We’re talking here of a collaborative research project in which UNC scientists along with colleagues in Barcelona have generated a complete map of the areas of the genome that control which genes are “turned on” or “off.”

The discovery, made in pancreatic islet cells, could open new avenues for understanding the genetic basis of type 2 diabetes and other common illnesses.

Since the completion of the human genome project, thousands of sequences associated with human illness have been discovered. But pinpointing which sequence variations are the true culprits has proven difficult.

That’s because the underlying sequence alphabet – the letters for the  nucleotides A, C, T, and G that form the bases of the genetic code – are only part of the story.  So it’s  not just the message, but the chromatin packaging of DNA that determine which genes are active and which are not.

Jason Lieb, Ph.D.

“Most of the human genome is uncharted territory – entire stretches of sequence with no clear function or purpose,” said Jason Lieb, Ph.D., associate professor of Biology at UNC and one of the senior authors on the study. “In fact, the majority of the DNA sequences associated with disease found thus far reside in the middle of nowhere. Here we have developed a map that can guide scientists to regions of the genome that do appear to be functionally relevant, instead of a dead end.”

The research, published online Jan. 31, 2010, in the journal Nature Genetics, presents the first high-resolution atlas of these regulatory elements in the most studied cell type for treatment and prevention of type II diabetes.

Using a new method developed in the Lieb laboratory called FAIRE-seq, Lieb and his colleagues isolated and sequenced a total of 80,000 open chromatin sites within pancreatic islet cells. They then compared these sites to those in non-islet cells to narrow the number down to 3,300 clusters of sites specific to this cell type.

Each cluster typically encompassed single genes that are active specifically in islet cells. Twenty of these genes are known to harbor gene variants associated with type II diabetes.

The researchers decided to continue their studies on the variant most strongly associated with the disease, a single nucleotide polymorphism – or SNP – occurring in the TCF7L2 gene. They found that the chromatin is more open in the presence of the high risk version of the gene (a T) than in the presence of the non-risk version (an A).

Further analysis demonstrated that the risk variant enhanced the activity of the gene, indicating that it may possess functional characteristics that could contribute to disease.

Lieb says his map is likely to help others within the diabetes research community identify new targets for understanding – and ultimately treating – the disease more effectively. But the approach is not limited to diabetes, or even pancreatic islet cells.

Lieb says he plans to use FAIRE-seq to chart the open chromatin regions present within other cells, such as the immune system’s lymphocytes.

Les Lang and Marla Vacek Broadfoot