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Characterization of Rabaptin-5 γ isoform

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Abstract

Rab GTPases are key regulators of intracellular membrane traffic acting through their effector molecules. Rabaptin-5 is a Rab5 effector in early endosome fusion and connects Rab5- and Rab4-positive membrane compartments owing to its ability to interact with Rab4 GTPase. Recent studies showed that Rabaptin-5 transcript is subjected to extensive alternative splicing, thus resulting in expression of Rabaptin-5 isoforms mostly bearing short deletions in the polypeptide chain. As interactions of a Rab GTPase with different effectors lead to different responses, functional characterization of Rabaptin-5 isoforms becomes an attractive issue. Indeed, it was shown that Rab GTPase effector properties of Rabaptin-5 and its α and δ isoforms are different. This work focused on another Rabaptin-5 isoform, Rabaptin-5γ. Despite its ability to interact with Rab5, endogenously produced Rabaptin-5γ was absent from early endosomes. Rather, it was found to be tightly associated with trans-Golgi network and partially localized to a Rab4-positive membrane compartment. The revealed intracellular localization of Rabaptin-5γ indicates that it is not involved in Rab5-driven events; rather, it functions in other membrane transport steps. Our study signifies the role of alternative splicing in determination of functional activities of Rab effector molecules.

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Abbreviations

3AT:

3-amino-(1,2,4)-triazole

EGF:

epidermal growth factor

EGFP:

enhanced green fluorescent protein

GAL4AD:

transcriptional activating domain of GAL4 transcription factor

GAL4BD:

GAL4 transcription factor DNA binding domain

GST:

glutathione-S-transferase

mAb:

monoclonal antibody

PBS:

phosphate buffered saline

PFA:

paraformaldehyde

PNS:

post-nuclear supernatant

TGN:

trans-Golgi network

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Author notes

  1. S. L. Kiselev

    Present address: Vavilov Institute of General Genetics, ul. Gubkina 3, 119911, Moscow, Russia

Authors and Affiliations

  1. Institute of Gene Biology, Russian Academy of Sciences, ul. Vavilova 34/5, 119334, Moscow, Russia

    E. V. Korobko, S. L. Kiselev & I. V. Korobko

Authors
  1. E. V. Korobko
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  2. S. L. Kiselev
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  3. I. V. Korobko
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Correspondence to I. V. Korobko.

Additional information

Published in Russian in Biokhimiya, 2014, Vol. 79, No. 9, pp. 1070–1078.

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Korobko, E.V., Kiselev, S.L. & Korobko, I.V. Characterization of Rabaptin-5 γ isoform. Biochemistry Moscow 79, 856–864 (2014). https://doi.org/10.1134/S000629791409003X

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  • DOI: https://doi.org/10.1134/S000629791409003X

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