TY - JOUR
T1 - A molecular sensor reveals differences in macromolecular crowding between the cytoplasm and nucleoplasm
AU - Murade, Chandrashekhar U.
AU - Shubeita, George T.
N1 - Funding Information:
This work was supported in part by the National Science Foundation Grants PHY-1505020 and PHY-1915119 to G.T.S. The authors acknowledge Ibtissem Nabti and Tomas Venit for assistance with experimental protocols and Piergiorgio Percipalle for useful discussions. The research was partially carried out using the Core Technology Platform resources at New York University Abu Dhabi.
Funding Information:
This work was supported in part by the National Science Foundation Grants PHY-1505020 and PHY-1915119 to G.T.S. The authors acknowledge Ibtissem Nabti and Tomas Venit for assistance with experimental protocols and Piergiorgio Percipalle for useful discussions. The research was partially carried out using the Core Technology Platform resources at New York University Abu Dhabi.
Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/7/26
Y1 - 2019/7/26
N2 - We describe a molecular sensor that reports, using fluorescence resonance energy transfer (FRET), on the degree of macromolecular crowding in different cellular compartments. The oligonucleotide-based sensor is sensitive to changes in the volume fraction of macromolecules over a wide range in vitro and, when introduced in cells, rapidly distributes and shows a striking contrast between the cytosol and the nucleus. This contrast can be modulated by osmotic stress or by using a number of drugs that alter chromatin organization within the nucleus. These findings suggest that the sensor can be used as a tool to probe chromosome organization. Further, our finding that the cell maintains different degrees of macromolecular crowding in the cytoplasm and nucleoplasm has implications on molecular mechanisms since crowding can alter protein conformations, binding rates, reaction kinetics, and therefore protein function.
AB - We describe a molecular sensor that reports, using fluorescence resonance energy transfer (FRET), on the degree of macromolecular crowding in different cellular compartments. The oligonucleotide-based sensor is sensitive to changes in the volume fraction of macromolecules over a wide range in vitro and, when introduced in cells, rapidly distributes and shows a striking contrast between the cytosol and the nucleus. This contrast can be modulated by osmotic stress or by using a number of drugs that alter chromatin organization within the nucleus. These findings suggest that the sensor can be used as a tool to probe chromosome organization. Further, our finding that the cell maintains different degrees of macromolecular crowding in the cytoplasm and nucleoplasm has implications on molecular mechanisms since crowding can alter protein conformations, binding rates, reaction kinetics, and therefore protein function.
KW - DNA-based sensor
KW - FRET
KW - chromatin organization
KW - macromolecular crowding
KW - molecular sensor
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U2 - 10.1021/acssensors.9b00569
DO - 10.1021/acssensors.9b00569
M3 - Article
C2 - 31250628
AN - SCOPUS:85070555463
SN - 2379-3694
VL - 4
SP - 1835
EP - 1843
JO - ACS Sensors
JF - ACS Sensors
IS - 7
ER -