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It is clear that multiple kinesins can

transport a cargo over much greater
distances than a single motor [13].
What isnt clear is the benefit of using
two types of kinesin with different
velocities. Mechanical competition
was first invoked to explain the
intraflagellar transport of Bardet-Biedl
syndrome proteins in C. elegans [11].
Intraflagellar transport particles are
also transported by two kinesins that
differ in velocity kinesin-II and
OSM-3. Pan et al. [11] found that the
faster OSM-3 exerts an assisting load
on the slower kinesin-II, which in turn
exerts a hindering load on OSM-3.
The result was an intermediate velocity
that could be explained by mechanical
competition. The proposal is that
independent control of two kinesins
gives the nematode a greater ability
to modulate intraflagellar transport
and thereby produce distinct types
of cilia [17].
Why would Xenopus use two
different chromokinesins to
produce the polar ejection force? The
kinesin-10, Xkid, dominates in the
mechanical competition and produces
severe chromosome alignment defects
when depleted from egg extracts [18].
Indeed, the complete absence of the
kinesin-4, Xklp1, only decreased
microtubule gliding velocities twofold.
Xklp1 participates, however, in the
early stages of mitotic spindle
assembly. It acts essentially as
a chromosome passenger protein,
binding to chromosomes in metaphase
and relocalizing to the spindle midzone
in anaphase [8]. A small contribution to
the polar ejection force may be the toll
that Xklp1 pays for its ride to the
midzone.

The emergent theme is that motor

coordination occurs through the
transmission of load. This is true for
situations involving a tug-of-war [19]
and situations described by Bieling and
colleagues [6]. Although there is good
evidence for regulatory molecules that
coordinate motor proteins in
Drosophila [4], the reconstitution of
multi-motor systems makes it clear
they are not necessary in vitro. The
transmission of assisting loads and
hindering loads between motor
proteins in an ensemble is sufficient to
coordinate them.
References
1. Howard, J. (2001). Mechanics of Motor Proteins
and the Cytoskeleton (Sunderland: Sinauer
Associates).
2. Wu, X., Bowers, B., Rao, K., Wei, Q., and
Hammer, J.A., 3rd (1998). Visualization of
melanosome dynamics within wild-type and
dilute melanocytes suggests a paradigm for
myosin V function In vivo. J. Cell Biol. 143,
18991918.
3. Morris, R.L., and Hollenbeck, P.J. (1993). The
regulation of bidirectional mitochondrial
transport is coordinated with axonal outgrowth.
J. Cell Sci. 104, 917927.
4. Welte, M.A., Gross, S.P., Postner, M.,
Block, S.M., and Wieschaus, E.F. (1998).
Developmental regulation of vesicle transport
in Drosophila embryos: forces and kinetics. Cell
92, 547557.
5. Snow, J.J., Ou, G., Gunnarson, A.L.,
Walker, M.R., Zhou, H.M., Brust-Mascher, I.,
and Scholey, J.M. (2004). Two anterograde
intraflagellar transport motors cooperate to
build sensory cilia on C. elegans neurons. Nat.
Cell Biol. 6, 11091113.
6. Bieling, P., Kronja, I., and Surrey, T. (2010).
Microtubule motility on reconstituted meiotic
chromatin. Curr. Biol. 20, 763769.
7. Rieder, C.L., and Salmon, E.D. (1994).
Motile kinetochores and polar ejection
forces dictate chromosome position on the
vertebrate mitotic spindle. J. Cell Biol. 124,
223233.
8. Vernos, I., Raats, J., Hirano, T., Heasman, J.,
Karsenti, E., and Wylie, C. (1995). Xklp1,
a chromosomal Xenopus kinesin-like
protein essential for spindle organization
and chromosome positioning. Cell 81,
117127.

Plant Cell Biology: How to Pattern

a Wall
How are the different secondary cell wall deposition patterns generated in the
vascular cells of plants? The use of a novel Arabidopsis mesophyll cell culture
that transdifferentiates into vascular cells shows a crucial role for a complex of
two microtubule-binding proteins.
Herman Hofte
Plant cells can adopt a remarkable
diversity of shapes, such as
star-shaped parenchyma cells, conical

epidermal cells, extremely elongated

and branched trichomes and tubular
water-conducting xylem cells
(Figure 1A). Local deposition and
remodeling of the primary cell wall is

9. Antonio, C., Ferby, I., Wilhelm, H., Jones, M.,

Karsenti, E., Nebreda, A.R., and Vernos, I.
(2000). Xkid, a chromokinesin required for
chromosome alignment on the metaphase
plate. Cell 102, 425435.
10. Howard, J., Hudspeth, A.J., and Vale, R.D.
(1989). Movement of microtubules by
single kinesin molecules. Nature 342,
154158.
11. Pan, X., Ou, G., Civelekoglu-Scholey, G.,
Blacque, O.E., Endres, N.F., Tao, L.,
Mogilner, A., Leroux, M.R., Vale, R.D., and
Scholey, J.M. (2006). Mechanism of transport of
IFT particles in C. elegans cilia by the concerted
action of kinesin-II and OSM-3 motors. J. Cell
Biol. 174, 10351045.
12. Larson, A.G., Landahl, E.C., and Rice, S.E.
(2009). Mechanism of cooperative behaviour in
systems of slow and fast molecular motors.
Phys. Chem. Chem. Phys. 11, 48904898.
13. Klumpp, S., and Lipowsky, R. (2005).
Cooperative cargo transport by several
molecular motors. Proc. Natl. Acad. Sci. USA
102, 1728417289.
14. Muller, M.J., Klumpp, S., and Lipowsky, R.
(2008). Tug-of-war as a cooperative mechanism
for bidirectional cargo transport by molecular
motors. Proc. Natl. Acad. Sci. USA 105,
46094614.
15. Grill, S.W., Kruse, K., and Julicher, F. (2005).
Theory of mitotic spindle oscillations. Phys.
Rev. Lett. 94, 108104.
16. Leduc, C., Ruhnow, F., Howard, J., and Diez, S.
(2007). Detection of fractional steps in cargo
movement by the collective operation of
kinesin-1 motors. Proc. Natl. Acad. Sci. USA
104, 1084710852.
17. Evans, J.E., Snow, J.J., Gunnarson, A.L.,
Ou, G., Stahlberg, H., McDonald, K.L., and
Scholey, J.M. (2006). Functional modulation
of IFT kinesins extends the sensory repertoire
of ciliated neurons in Caenorhabditis elegans.
J. Cell Biol. 172, 663669.
18. Funabiki, H., and Murray, A.W. (2000). The
Xenopus chromokinesin Xkid is essential for
metaphase chromosome alignment and must
be degraded to allow anaphase chromosome
movement. Cell 102, 411424.
19. Soppina, V., Rai, A.K., Ramaiya, A.J., Barak, P.,
and Mallik, R. (2009). Tug-ofwar between
dissimilar teams of microtubule motors
regulates transport and fission of endosomes.
Proc. Natl. Acad. Sci. USA 106, 1938119386.

Department of Biology, McGill University,

Montreal, Quebec H3A 1B1, Canada.
E-mail: gary.brouhard@mcgill.ca
DOI: 10.1016/j.cub.2010.03.034

responsible for the shape of the

growing cell. Deposition of a secondary
wall after growth cessation determines
the mechanical properties of the
mature cell. For instance, in xylem
cells, resistance against the negative
pressure of the water stream is
conditioned by cell wall reinforcement
patterns, which are spiral or annular
in protoxylem (i.e., xylem produced
from the meristematic cells in
the procambium) and reticulate or
pitted in metaxylem (develops
after the protoxylem) (Figure 1B).
A long-standing question in plant

Dispatch
R451

biology is how these specific cell wall

patterns are specified. The
development of Zinnia elegans [1]
mesophyll cell suspensions, and
later Arabidopsis cell lines [2], that
are hormonally triggered to
transdifferentiate into vascular cells,
referred to as tracheary elements, have
been instrumental for the study of the
differentiation of this specialized cell
type. Mature tracheary elements are
dead cells generated in four steps: cell
expansion, secondary wall deposition,
lignification and cell death.
Transcriptional profiling in these cells
combined with mutant analysis in
Arabidopsis has provided insights into
the transcription regulation network
underlying these developmental events
[3]. For instance, the NAM, ATAF and
CUC (NAC) transcription factors VND7
and VND6 are involved in the formation
of protoxylem and metaxylem,
respectively. Interestingly, ectopic
expression of these transcription
factors can trigger transdifferentiation
of mesophyll or epidermis cells into the
respective cell types, indicating that
these transcription factors regulate,
directly or indirectly, sets of genes
controlling the deposition of the
corresponding cell wall patterns [2]. A
number of downstream genes involved
in cellulose or lignin synthesis have
been identified; however, it is
not known how the specific cell wall
deposition patterns are generated.
The walls of tracheary elements
consist of an extremely resistant
composite material, reminiscent of
reinforced concrete, in which tensile
stiffness is provided by oriented
cellulose microfibrils and compression
resistance by the lignified matrix.
Cellulose is the major organizer of cell
wall architecture. It is synthesized by
25-nanometer complexes, which
contain a large number of cellulose
synthase catalytic subunits that
extrude -1,4-linked glucan chains,
which assemble into crystalline
microfibrils. Cortical microtubules play
a key role in this process: they target
the insertion of cellulose synthase
complexes to specific plasma
membrane domains [4,5] and
subsequently guide their trajectories in
the plasma membrane while they spin
out the cellulose [6]. This cell-wall
patterning mechanism can be
generalized to many plant cell types,
including tracheary elements [7]. The
next question is how the microtubules
are organized?

Current Biology

Figure 1. The cell wall conditions the shape and mechanical properties of plant cells.
(A) Plant cells can adopt a large diversity of shapes. Examples shown are star-shaped parenchyma cells, conical epidermal cells, extremely elongated and branched trichomes or tubular
water-conducting xylem cells (reproduced with permission from the American Society of Plant
Biologists). (B) Scanning electron micrograph of longitudinal section through vascular tissue in
Arabidopsis. Cell wall deposition patterns in xylem vessel cells: spiral and annular in protoxylem cells (arrowhead); reticulate and pitted in metaxylem cells (arrow). Image courtesy of
Preeti Dahiya, Kim Findlay, and Keith Roberts.

The presence of arrays of mainly

parallel interphase cortical
microtubules involved in cell wall
deposition distinguishes plant cells
from other eukaryotic cells.
Microtubules are also highly dynamic in
these arrays and can rapidly change
orientation or form bundles. For
instance, in cells approaching mitosis,
cortical microtubules bunch up and
form the so-called preprophase band
that encircles the cell at the position of
the future crosswall. Similarly, during
tracheary element differentiation,
microtubules form bundles beneath
the future cell wall thickenings. In
a recent issue of Current Biology,
Pesquet et al. [8] identified a key actor
in the organization of microtubules
during tracheary element
differentiation. The authors first
selected a new Arabidopsis cell culture
in which transdifferentiation into
tracheary elements can be triggered at
high efficiency (up to 40%) by the
addition of auxin, cytokinin and
brassinosteroid hormones. Using an
elegant time-lapse imaging method,
the differentiation sequence was
shown to involve: cellulose deposition
for 68 hours, a lag time of 2 hours,
nuclear degradation for 10 minutes,
and finally protoplast shrinkage and
cell wall lignification for up to 4 hours.
Interestingly, the lignification appeared
to occur postmortem. It will be
interesting to see whether this
lignification reflects an oxidative
burst triggering the polymerization

of monolignols previously secreted

into the wall or the release of
monolignols from the dying cell. Next,
they used transcript profiling data from
Zinnia cell suspensions undergoing
tracheary element differentiation to
identify, among 200 known
microtubule-associated proteins
(MAPs), only one tracheary element
specific protein that was upregulated
during differentiation, AtMAP70-5.
This protein was previously shown
to stabilize microtubules in vitro. Yeast
two-hybrid analysis and reciprocal
co-immunoprecipitation showed that
AtMAP70-5 interacts with AtMAP70-1,
a related isoform that is constitutively
expressed in tracheary elements. Live
cell imaging using an a-tubulinGFP
fusion protein showed that microtubule
bundles precisely underlie the cell wall
thickenings. In contrast, GFP-fused
AtMAP70-1 or AtMAP70-5 labeled
a sub-class of microtubules that
straddle the microtubule-covered
plasma membrane area beneath the
wall thickening (Figure 2A), suggesting
that the AtMAP70s play a role in
patterning. This was confirmed by
showing that it was possible to
manipulate the cell wall pattern by
modulating the amount of either
AtMAP70. Simultaneous
overexpression of AtMAP70-1 or -5
increased the percentage of spiral
walls and reduced the percentage of
pitted walls (Figure 2B). Silencing the
AtMAP70-1 or -5 instead increased
the percentage of pitted walls. These

Current Biology Vol 20 No 10

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Control tracheary element

cell line

MAP70 RNAi tracheary element

cell line

deposition patterns during wood

formation can be controlled by
microtubule-binding proteins.
References

B
More

Less
MAP70

MAP70-labelled
microtubules

Plasma membrane

Other microtubules

Cell wall
Current Biology

Figure 2. A schematic model of how MAP70 proteins might position cortical microtubules and
control cell wall patterning in developing tracheary elements.
(A) In control tracheary elements, microtubules (green) form a U-shaped mould while AtMAP70-5
and -1 (light purple) decorate those microtubules flanking the thickenings adjacent to the primary
cell wall (purple). RNAi silencing causes the separation of the thickening and its associated
microtubules from the primary cell wall. (B) Schematic representation of how the secondary
thickening pattern might be regulated by modulating the amount of AtMAP70-5 and -1. Larger
amounts of AtMAP70 increase the spacing between thickenings, resulting in an increased
percentage of spiral patterns among the tracheary elements. Reduction of the amount of
AtMAP70 reduces the spacing between thickenings, leaving only unthickened pits within the
secondary wall.

results indicate that the abundance of

the AtMAP70-1AtMAP70-5 complex
in the cell is sufficient to explain the
different cell wall patterns. It remains
unclear, however, whether the relative
abundance of the AtMAP70s also
explains the cell wall patterns in
wild-type proto- and metaxylem cells
and whether the transcription factors
VND7 and VND6 differentially regulate
their expression levels. Another, rather
bizarre finding was that, in the silenced
lines for either AtMAP70, about half of
the thickenings were detached from the
surface and formed internalized
strands consisting of cell wall material
surrounded by plasma membrane and
microtubules. The authors propose
that AtMAP70-decorated microtubules
may define the boundary between
the thickenings and the primary cell
wall. In the absence of such a boundary
in the RNAi lines, the thickening with its
associated plasma membrane and
microtubules can disconnect from
the rest of the plasma membrane
(Figure 2A). Live cell imaging during

tracheary element differentiation in

the silenced lines should provide
insights into the mechanism of the
internalization of the thickenings.
In conclusion, this study provides
new insights into how cell wall

1. Fukuda, H., and Komamine, A. (1980).

Establishment of an experimental system for
the study of tracheary element differentiation
from single cells isolated from the mesophyll
of Zinnia elegans. Plant Physiol. 65, 5760.
2. Kubo, M., Udagawa, M., Nishikubo, N.,
Horiguchi, G., Yamaguchi, M., Ito, J., Mimura, T.,
Fukuda, H., and Demura, T. (2005). Transcription
switches for protoxylem and metaxylem vessel
formation. Genes Dev. 19, 18551860.
3. Demura, T., and Fukuda, H. (2007).
Transcriptional regulation in wood formation.
Trends Plant Sci. 12, 6470.
4. Crowell, E.F., Bischoff, V., Desprez, T.,
Rolland, A., Stierhof, Y.D., Schumacher, K.,
Gonneau, M., Hofte, H., and Vernhettes, S.
(2009). Pausing of Golgi bodies on microtubules
regulates secretion of cellulose synthase
complexes in Arabidopsis. Plant Cell 21,
11411154.
5. Gutierrez, R., Lindeboom, J.J., Paredez, A.R.,
Emons, A.M., and Ehrhardt, D.W. (2009).
Arabidopsis cortical microtubules position
cellulose synthase delivery to the plasma
membrane and interact with cellulose synthase
trafficking compartments. Nat. Cell Biol. 11,
797806.
6. Paredez, A.R., Persson, S., Ehrhardt, D.W., and
Somerville, C.R. (2008). Genetic evidence that
cellulose synthase activity influences
microtubule cortical array organization. Plant
Physiol. 147, 17231734.
7. Oda, Y., Mimura, T., and Hasezawa, S. (2005).
Regulation of secondary cell wall development
by cortical microtubules during tracheary
element differentiation in Arabidopsis cell
suspensions. Plant Physiol. 137, 10271036.
8. Pesquet, E., Korolev, A.V., Calder, G., and
Lloyd, C.W. (2010). The microtubule-associated
protein AtMAP70-5 regulates secondary wall
patterning in Arabidopsis wood cells.
Curr. Biol. 20, 744749.

Institut Jean-Pierre Bourgin, UMR1318

INRA-AgroParisTech, INRA Centre de
Versailles-Grignon, Route de St-Cyr (RD10),
78026 Versailles Cedex, France.
E-mail: herman.hofte@versailles.inra.fr

DOI: 10.1016/j.cub.2010.03.046

Cellular Homeostasis: Coping with ER

Overload During an Immune
Response
Host cells secrete antimicrobial proteins en masse to counter extracellular
pathogens, placing a strain on the endoplasmic reticulum. The interplay
between defence and cellular homeostasis has now been dissected genetically
in Caenorhabditis elegans.
Jonathan J. Ewbank1,2,3,*
and Nathalie Pujol1,2,3
Immunologists are an isolated breed.
Their work is shrouded in mystery, with

impenetrable codes and such

a specialized vocabulary that it takes
the uninitiated years to be fully versant
in the discipline. In part, this is
a consequence of a long-standing