Project Title

Cracking the Code for Intranuclear Trafficking: Unlocking Bruno Protein

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Faculty Sponsor Name

Anton Bryantsev

Disciplines

Biology | Biotechnology | Cell and Developmental Biology | Cell Biology | Molecular Genetics

Abstract (300 words maximum)

Despite the lack of internal membranes, the cell nucleus is neatly organized into various kinds of defined territories with sharp boundaries, termed nuclear domains (NDs). Each ND type accumulates specific nuclear proteins and maintaining the harmony of this protein traffic is vital for the proper functions of nuclear regulatory machinery. Studying the code of intranuclear trafficking and sorting is posed to enable better control over the nuclear functions for research and biotechnological needs.

Our research model was based on B-bodies, the recently discovered NDs from the Drosophila muscles. One of the resident proteins of B-bodies, Bruno, is composed of three RNA-binding motifs (RRMs) and two internally disordered regions (IDRs). We applied experimental genetics and mutagenesis to isolate the essential protein sequence required for accumulation into B-bodies. All mutant sequences were tagged with green fluorescent protein (GFP), and their accumulation in B-bodies was evaluated by immunofluorescence.

After analyzing 9 mutants, we have identified the minimal sequence for trafficking to B-bodies, which consists of two RRM and one IDR domains. Our data suggest that RNA binding ability is key to the specificity in ND trafficking. In the future, we could use this knowledge as a blueprint for modifying nuclear proteins that can safely navigate intranuclear traffic by adding a B-body tag to drive excess of proteins of interest to B-bodies

Project Type

Oral Presentation (15-min time slots)

How will this be presented?

Yes, in person

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Cracking the Code for Intranuclear Trafficking: Unlocking Bruno Protein

Despite the lack of internal membranes, the cell nucleus is neatly organized into various kinds of defined territories with sharp boundaries, termed nuclear domains (NDs). Each ND type accumulates specific nuclear proteins and maintaining the harmony of this protein traffic is vital for the proper functions of nuclear regulatory machinery. Studying the code of intranuclear trafficking and sorting is posed to enable better control over the nuclear functions for research and biotechnological needs.

Our research model was based on B-bodies, the recently discovered NDs from the Drosophila muscles. One of the resident proteins of B-bodies, Bruno, is composed of three RNA-binding motifs (RRMs) and two internally disordered regions (IDRs). We applied experimental genetics and mutagenesis to isolate the essential protein sequence required for accumulation into B-bodies. All mutant sequences were tagged with green fluorescent protein (GFP), and their accumulation in B-bodies was evaluated by immunofluorescence.

After analyzing 9 mutants, we have identified the minimal sequence for trafficking to B-bodies, which consists of two RRM and one IDR domains. Our data suggest that RNA binding ability is key to the specificity in ND trafficking. In the future, we could use this knowledge as a blueprint for modifying nuclear proteins that can safely navigate intranuclear traffic by adding a B-body tag to drive excess of proteins of interest to B-bodies

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