School of Science Division of Life Science 16 Functional Roles of Arfpr1 in Regulating Export of Cargo Proteins out of the Trans Golgi Network Supervisor: GUO Yusong / LIFS Student: LAI Chak Long / BCB-IRE Course: UROP 1100, Summer At the trans-Golgi network, small GTPases of the Arf family recruit cytosolic cargo adaptors to the membrane. One of the Arf family proteins, Arf-related protein 1 (Arfrp1), is responsible for the membrane recruitment of cytosolic adaptor complex 1 (AP-1), the major cargo adaptor. While the guanine nucleotide exchange factor (GEF) for Arfrp1 remains unknown, Golgi trafficking protein, Sys1, is reported to be critical for recruiting Arfrp1 to the Golgi. Therefore, we hypothesised that Sys1 is the GEF for Arfrp1. To test this hypothesis, GST pull-down and co-IP experiments were conducted to test whether Arfrp1-AP-1 binding is Sys1-dependent. Additionally, co-IP and immunofluorescence experiments were conducted to test the roles of Sys1 N- and C-terminals in the membrane recruitment of Arfrp1. Functional Roles of Arfpr1 in Regulating Export of Cargo Proteins out of the Trans Golgi Network Supervisor: GUO Yusong / LIFS Student: XU Tongle / BCB Course: UROP 1100, Summer The Sonic Hedgehog (Shh) signaling pathway is critical for embryonic development and various cellular processes. Aberrant activation of this pathway is linked to multiple cancers. In this study, we reselected nanobodies targeting ShhN from a selection yeast pool and identified four new nanobody sequences. Additionally, we successfully modified the plasmids of Nanobody 1-5, inserting three base pairs in its conversed coding sequence by Quickchange experiments. Sequencing results confirmed the restoration of the complete sequence. Further experiments are needed to assess whether the modified nanobody exhibits improved efficacy. These findings could advance our understanding of the Shh pathway and aid in developing targeted cancer therapies. Biochemical Characterization of Histone Variants and Post-translationally Modified Nucleosomes Supervisor: ISHIBASHI Toyotaka / LIFS Student: HARIMOTO Yuken / BCB Course: UROP 3100, Spring While histones proteins are widely known for their roles in organizing DNA, they have also been recognized to have antimicrobial properties and being deployed by organisms as a defence mechanism against various microorganisms. In this UROP project, we investigate those properties of histones and subsequent nucleosomes by testing the antimicrobial properties in an assay against Escherichia coli (E.coli) and Saccharomyces cerevisiae (S.cerevisiae). The results show that throughout different histone proteins, H2A and H2B FWT were the most effective in antimicrobial activities against E.Coli while H2A and H2B K works the best against S.cerevisiae. In an assay against microbials using nucleosomes, unloaded histone octamers performed the best against both E.Coli and S.cerevisiae compared to canonical nucleosomes and nucleosome incorporating variants.
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