An ezrin-rich, rigid uropod-like structure directs movement of

amoeboid blebbing cells

Anna Lorentzen, Jeffrey Bamber, Amine Sadok, Ilan Elson-Schwab and Christopher J. Marshall.

Tamara Heredia

27/11/2014

Blebbing

Fig. 1 Schematic of a bleb formation. (Fackler et al, 2008)

Single migrating cells

Mesenchymal-type elongated mode

Rounded mode of movement or

Ameboid mode

Requires degradation of the ECM by

proteases

Is less dependent on proteases. Cells move

by membrane blebbing or by squeezing
through pores in the ECM.

Requires low actomyosin contractility

High degree of actomyosin contractility

Elongated morphology

Rounded morphology
(Amit Pathak et al, 2010)

Fig 2. Illustration of the establishment of two poles in migrating leukocytes, adhesion molecules
and cytoskeletal elements. (Francisco Sanchez-Madrid1 et al, 1999)

Note: Uropod also contains protein kinase A (PKA) and phosphatidylinositol (4,5)bisphosphate [PtdIns(4,5)P2]

Uropods contain:

Membrane receptors

Signalling molecules

Organelles

Integrins

Ezrin-radixin-moesin
(ERM) family proteins

Golgi complex

Intercellular adhesion
molecules (ICAMs)

Protein kinase A (PKA)

Microtubule organising
centre (MTOC)

Hyaluronic acid receptor

CD44

Phosphatidylinositol (4,5)Bisphosphate
[PtdIns(4,5)P2]

The uropod contains:

P-selectin glycoprotein
ligand-1 (PSGL-1)

Nonadhesive amoeboid movement

Characterised by the formation of
membrane blebs

It has been observed in Dictyostelium and

cancer cells.

Cells with blebbing amoeboid movement

lack apparent polarisation and have high
actomyosin contractility .

Fig. 3 (A) Amoeboid cell crawling on a twodimensional surface. Lamellipodium

formation at the front determines the
direction of movement. The front of the
cell adheres to the substrate in focal
adhesions through integrins. Retraction
of the uropod at the rear creates the force
that moves the cell forward. (B)
Amoeboid cell squeezing through pores
in a three-dimensional matrix. A
membrane bleb grows into a gap in the
ECM. (C )Movement by cells forming
multiple small blebs. High contractility
of the cell creates pressure that is
released in membrane blebs.

In this research:
1. It is investigated the polarisation in blebbing amoeboid A375 melanoma cells in order to

understand cell movement driven by multiple blebs.

A375 cells: human melanoma cell with invasive and metastatic properties. A375 melanoma cells
form multiple blebs over the membrane and would therefore not be able to move in one determined
direction.
A375P means that the cell line contain low concentration of A375, A375M-medium and A375SMhigh concentration.
2. It is shown that A375 cells form an ezrin-rich uropod-like structure (to which the autors call the
ERULS) at their cell rear, and that this is required for invasion into a three-dimensional matrix.
3. It is presented a model explaining how cells can move without a defined front and only a defined
rear.

RESULTS
1. Polarisation of blebbing amoeboid cells

[PtdIns(4,5)P
2]

2. Ezrin accumulates at the cell rear

3. ROCK and novel PKC isoforms regulate the ERULS

4. The Golgi is located in front of the nucleus in blebbing cells

5. The ERULS is an inherent polarisation structure of rounded
blebbing cells

6. The ERULS has an active role in blebbing amoeboid cell

invasion

7. Blebbing is decreased at the ERULS

Model
.

Summary

It was investigated how melanoma cells with a round morphology and many small blebs over their
surface can move without displaying morphological features of a front or a back.
It was shown that blebbing melanoma cells form a uropod-like structure (ERULS) at the rear that is
enriched in ezrin, b1-integrin, actin, MLC and PtdIns(4,5)P2.
It was shown that this structure is a feature of polarised round blebbing cells and is not a
consequence of cell movement.
It was shown that the ERULS is enriched in the phosphorylated active form of ezrin.
It was identified the Golgi alignment in front of the nucleus in blebbing melanoma cells, which is
the opposite of the alignment in migrating leukocytes.
Also it was proposed a model explaining how cells can move without a defined front and only a
defined rear.

References

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