Functional assessment of the V390F mutation in the CCT delta subunit of chaperonin containing tailless complex polypeptide 1

• The chaperonin containing tailless complex polypeptide 1 (CCT) is a multi-subunit molecular chaperone. It is found in the cytoplasm of all eukaryotic cells, where the oligomeric form plays an essential role in the folding of predominantly the cytoskeletal proteins actin and tubulin. Both the CCT oligomer and monomeric subunits also display functions that extend beyond folding, which are often associated with microtubules and actin filaments. Here, we assess the functional significance of the CCT delta V390F mutation, reported in several cancer cell lines. Upon transfection into B16F1 mouse melanoma cells, GFP-CCT delta(V390F) incorporates into the CCT oligomer more readily than GFP-CCT delta. Furthermore, unlike GFP-CCT delta, GFP-CCT delta(V390F) does not interact with the dynactin complex component, p150(Glued). As CCT delta has previously been implicated in altered migration in wound healing assays, we assessed the behaviour of GFP-CCT delta(V390F) and other mutants of CCT delta, previously used to assess functional interactions with p150(Glued), in chemotaxis assays. We developed the assay system to incorporate a layer of the inert hydrogel GrowDex (R) to provide a 3D matrix for chemotaxis assessment and found subtle differences in the migration of B16F1 cells, depending on the presence of the hydrogel.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)

Nyckelord

Molecular chaperone

Cancer cell line

Chemotaxis

GrowDex (R)

cytosolic chaperonin

protein gelsolin

expression

tubulin

cells

Cell Biology

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