The nuts and bolts of copying DNA, $12 million, five-year grant from the National Institute of General Medical Science

Researchers: John Lis, professor of molecular biology and genetics in the College of Agriculture and Life Sciences (lead PI); Steven Josefowicz, assistant professor of pathology and laboratory medicine at Weill Cornell Medicine; Effie Apostolou, associate professor of molecular biology in medicine at Weill Cornell Medicine; Haiyuan Yu, professor of biological statistics and computational biology in CALS; and Abdullah Ozer, a research associate in Lis’ lab.

A better, more complete understanding of the transcription cycle – where DNA is copied into RNA, which is translated into proteins – and its regulation could lead to new targets for therapies for cancer, neurological and developmental disorders, autoimmune, metabolic and cardiovascular diseases related to misregulation of transcription.

The team will use a bevy of state-of-the-art technologies to investigate large macromolecular complexes involved in transcription and its regulation. They developed an RNA aptamer – RNA selected to tightly bind to a green fluorescent protein ­­– for tagging large complexes. The group will focus initially on RNA Polymerase II, the main machine that transcribes mRNA encoding genes.

“These kinds of collaborations are so fulfilling for junior faculty like me, who can learn from him rather than having to reinvent the wheel. It means having so much more fun and getting to work with other passionate scientists.”

Steven Josefowicz

“We want to tag a subunit of the polymerase as one of the macromolecular complexes we’re interested in and be able to then track it as it moves through its various regulated steps in the transcription cycle,” Lis said.

In this way, the researchers can detect complexes and their associated protein partners and potential regulators.

A special mass spectrometry machine will allow them to dissect the composition of a complex at various stages of the cycle; a cryo-electron microscope will provide a high-resolution structure of a complex; cross-linking reagents will capture proteins loosely associated in the cycle; and new software will aid analysis.

The team expertise will allow them to study native complexes during transcription, “so we can see it as it really is in the cell,” Josefowicz said.

Lis and colleagues received a $100,000 academic integration seed grant in late 2019.

Also, the project teamed senior researchers like Lis with junior faculty, such as Josefowicz, who face challenges of setting up a lab from scratch and maximizing the potential reach of their nascent research programs.

“John’s a giant in this field,” Josefowicz said. “These kinds of collaborations are so fulfilling for junior faculty like me, who can learn from him rather than having to reinvent the wheel. It means having so much more fun and getting to work with other passionate scientists.”