Deciphering the cellular functions of the Op18/Stathmin family of microtubule-regulators by plasma membrane-targeted localization.
The Op18/stathmin family of microtubule regulators includes the ubiquitous cytosolic Op18/stathmin (Op18) and the neuronal, primarily Golgi-associated proteins SCG10 and RB3, which all form ternary complexes with two head-to-tail-aligned tubulin heterodimers. To understand the physiological significance of previously observed differences in ternary complex stability, we have fused each of the heterodimer-binding regions of these three proteins with the CD2 cell surface protein to generate confined plasma membrane localization of the resulting CD2 chimeras. Herein, we show that, in contrast to constitutively active CD2-Op18-tetraA, both the CD2-SCG10 and CD2-RB3 chimeras sequestered tubulin at the plasma membrane, which results in >35% reduction of cytosolic tubulin heterodimer levels and consequent delayed formation of mitotic spindles. However, all three CD2 chimeras, including the tubulin sequestration-incompetent CD2-Op18-tetraA, destabilize interphase microtubules. Given that microtubules are in extensive contact with the plasma membrane during interphase, but not during mitosis, these findings indicate that Op18-like proteins have the potential to destabilize microtubules by both sequestration and direct interaction with microtubules. However, the differences in tubulin binding observed in cells also indicate conceptual differences between the functions of low-abundance neural family members, which will accumulate tubulin at specific cellular compartments, and the abundant cytosolic Op18 protein, which will not.
Holmfeldt
Per
P
Department of Molecular Biology University of Umeå, SE-901 87 Umeå, Sweden.
Brannstrom
Kristoffer
K
Stenmark
Sonja
S
Gullberg
Martin
M
D018801:CD2 Antigens; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D019281:Dimerization; D005455:Fluorescent Antibody Technique; D006801:Humans; D007399:Interphase / Q000502:physiology; D020014:K562 Cells; D008565:Membrane Proteins; D008868:Microtubule Proteins; D008870:Microtubules / Q000378:metabolism* / Q000502:physiology; D008938:Mitosis / Q000502:physiology; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011485:Protein Binding; D017433:Protein Structure, Secondary / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins; D008941:Spindle Apparatus / Q000378:metabolism / Q000502:physiology; D050777:Stathmin; D020349:Surface Plasmon Resonance; D014404:Tubulin / Q000378:metabolism* / Q000502:physiology
D018801:CD2 Antigens; D008565:Membrane Proteins; D008868:Microtubule Proteins; D009414:Nerve Growth Factors; D010750:Phosphoproteins; D011994:Recombinant Proteins; C493995:STMN1 protein, human; C063314:STMN2 protein, human; C491795:STMN4 protein, human; D050777:Stathmin; D014404:Tubulin