Multidirectional Activity Control of Cellular Processes by a Versatile Chemo-optogenetic Approach

Article abstract of DOI:10.1002/anie.201806976

The spatiotemporal dynamics of proteins or organelles plays a vital role in controlling diverse cellular processes. However, acute control of activity at distinct locations within a cell is challenging. A versatile multidirectional activity control (MAC) approach is presented, which employs a photoactivatable system that may be dimerized upon chemical inducement. The system comprises second-generation SLF*-TMP (S*T) and photocaged NvocTMP-Cl dimerizers; where, SLF*-TMP features a synthetic ligand of the FKBP(F36V) binding protein, Nvoc is a caging group, and TMP is the antibiotic trimethoprim. Two MAC strategies are demonstrated to spatiotemporally control cellular signaling and intracellular cargo transport. The novel platform enables tunable, reversible, and rapid control of activity at multiple compartments in living cells.

Full text of DOI:10.1002/anie.201806976

A Journal of the Gesellschaft Deutscher Chemiker
Angewandte
Chemie
International Edition
www.angewandte.org
Accepted Article
Title: Multi-directional activity control (MAC) of cellular processes
enabled by a versatile chemo-optogenetic approach
Authors: Yaowen Wu, Xi Chen, Muthukumaran Venkatachalapathy,
and Leif Dehmelt
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: Angew. Chem. Int. Ed. 10.1002/anie.201806976
Angew. Chem. 10.1002/ange.201806976
Link to VoR: http://dx.doi.org/10.1002/anie.201806976
http://dx.doi.org/10.1002/ange.201806976

10.1002/anie.201806976
Angewandte Chemie International Edition
COMMUNICATION
Multi-directional activity control (MAC) of cellular processes
enabled by a versatile chemo-optogenetic approach
Xi Chen,[a,b] Muthukumaran Venkatachalapathy,[b,d] Leif Dehmelt,[b,d] Yao-Wen Wu*[a,b,c]
Abstract: The spatio-temporal dynamics of proteins or organelles
plays a vital role in controlling diverse cellular processes. However,
acute control of activity at distinct locations within a cell is challenging.
Here we present a versatile multi-directional activity control (MAC)
To unravel the complex and dynamic nature of such systems
that orchestrate cell function in space and time, acute
perturbations with high spatial and temporal resolution are
required. Conventional genetic manipulations, such as protein
overexpression, or siRNA-mediated knockdown, are limited as
they act very slowly on the entire cell. Chemically induced
dimerization (CID) is a powerful tool to modulate protein function
by bringing two proteins in close proximity to control their function
and to perturb associated cellular processes.[7] Optogenetic and
optochemical approaches, such as light-induced dimerization
confer superior spatiotemporal control to such perturbations.[8]
approach using
a
photoactivatable, dual-chemically induced
dimerization (pdCID) system by combining the second generation
SLF*-TMP (S*T) and photo-caged NvocTMP-Cl dimerizers. We
demonstrate two MAC strategies to spatiotemporally control cellular
signaling and intracellular cargo transport. We thereby provide a novel
platform that enables tunable, reversible, and rapid control of activity
at multiple compartments in living cells.
The spatio-temporal organization of cellular processes is
often controlled by the subcellular distribution of molecules
or organelles. As individual proteins can perform different
functions depending on their local environment, or when
tethered to different effectors, subcellular localization is
essential for the functional diversity of proteins.[1]
A
growing number of so called “moonlighting proteins” have
been identified, which play distinct roles when located at
different subcellular regions.[2] For instance, at the plasma
membrane, the Wnt-signaling protein β-catenin controls
cell-cell adhesion, while it also can regulate gene
transcription if it is localized in the nucleus.[3] Aberrant
localizations of proteins have been implicated in
pathogenesis of diverse human diseases and the
modulation of subcellular protein localization has emerged
as a target for therapeutic intervention.[4] Recent evidence
suggests a broad role of organelle distribution in the
spatial organization of several cellular processes,
including cell signaling, cell polarization and neurite
outgrowth.[5] For example, the bidirectional transport along axonal
microtubules plays a central role in the proper subcellular
distribution of cellular cargos and its misregulation is thought to
play an important role in neurodegenerative diseases.[6]
Figure 1. A) Previously developed systems only enable a single layer of activity control
of cellular processes; B) the MAC approach enables multi-directional activity control of
protein function and complicated cellular processes; the newly introduced 2nd
generation SLF*-TMP dimerizer (C) and the photo-caged NvocTMP-Cl dimerizer (D)
used in the pdCID system.
In the past, such approaches were successfully applied to
perform a single-layer of activity control, such as turn-on/off by
targeting a protein to a specific cellular compartment to study
relatively simple cellular processes (Figure 1A). However, the
analysis of more complex processes that involve cycling,
trafficking or shuttling of signal molecules between different cell
compartments, such as growth factor or Wnt signaling, could
benefit from multiple layers of activity control, e.g. by subsequent
targeting of proteins to distinct locations within a single cell (Figure
1B).[9] Herein we report a versatile photoactivatable, dual-
chemically induced dimerization (pdCID) system to enable acute,
switchable, and multi-directional activity control (MAC) in living
cells.
[a]
[b]
[c]
[d]
Dr. X. Chen, Prof. Dr. Y. W. Wu
Chemical Genomics Centre of the Max Planck Society
Otto-Hahn-Str. 15, 44227 Dortmund, Germany
E-mail: yaowen.wu@mpi-dortmund.mpg.de
Dr. X. Chen, Dr. M. Venkatachalapathy, Dr. L. Dehmelt, Prof. Dr. Y.
W. Wu
Max Planck Institute of Molecular Physiology
Otto-Hahn-Str. 11, 44227 Dortmund, Germany
Prof. Dr. Y. W. Wu
Department of Chemistry,
Umeå University, 90187 Umeå, Sweden
E-mail: yaowen.wu@umu.se
Dr. M. Venkatachalapathy, Dr. L. Dehmelt
Fakultät für Chemie und Chemische Biologie,
Technische Universität Dortmund
This system combines either the newly introduced second
generation SLF*-TMP (S*T) dimerizer or the first generation SLF'-
TMP (ST)[10] with the NvocTMP-Cl photocaged dimerizer,[11]
which were recently established in our lab (Figure 1, S1).
NvocTMP-Cl consists of caged trimethoprim (TMP), a HaloTag
ligand (chlorohexyl group) and a polyethylene glycol (PEG) linker.
TMP selectively binds to E.coli dihydrofolate reductase (eDHFR)
with a Kd of 1 nM and the chlorohexyl moiety can form a covalent
bond with a bacterial alkyl dehalogenase mutant (HaloTag).
Emil-Figge-Straße 50, 44227 Dortmund, Germany
Supporting information for this article is given via a link at the end of
the document
This article is protected by copyright. All rights reserved.

10.1002/anie.201806976
Angewandte Chemie International Edition
COMMUNICATION
NvocTMP-Cl can covalently pre-localize to a protein fused with a
HaloTag. The dimerization between HaloTag and eDHFR does
not occur until the nitroveratroyloxycarbonyl (Nvoc) caging group
is removed by illumination using a 405 nm light pulse (a process
called photolysis) (Figure S1). On the other hand, SLF*-TMP and
SLF'-TMP feature a synthetic ligand of FKBP(F36V) (SLF') and a
TMP moiety. SLF*-TMP (S*T) is an optically pure diastereomer
while SLF'-TMP (ST) is a diastereomeric mixture. SLF*/SLF' binds
in a high affinity (subnanomolar) to the FKBP(F36V) mutant
(FKBP') with a 1000-fold selectivity over wild-type FKBP. Both
dimerizers induce dimerization between FKBP'- and eDHFR-
fused proteins with nanomolar affinity (Figure S1F).[10] The
NvocTMP-Cl and S*T/ST-induced dimerization is stable and
essentially irreversible even after wash-out, but can be disrupted
by the competitor TMP to make both systems reversible (Figure
S1, S2, S3). The NvocTMP-Cl system is tunable by illumination
doses (Figure S4) and the induced dimerization is very rapid (t1/2
= 0.55 - 1.69 s) (Figure S5). Among many available dimerizer
systems, the combination of two is challenging and problematic,
because four dimerization modules are required and two
individual dimerization processes are difficult to connect with each
other. Herein, because the NvocTMP-Cl system shares a
common eDHFR dimerizing module with the S*T (or ST) system,
it is possible to combine these modular systems to enable multi-
directional control over protein or organelle activity in a single cell.
A major challenge in all dimerization systems is basal activity.
Even in the absence of dimerization, the protein of interest (POI)
can still reach the site of activation by diffusion through the cytosol,
resulting in background activity[7b-d]. A previous approach to
reduce background activity utilized a Golgi-binding fusion protein
to sequester the POI away from its target localization at the
plasma membrane. Upon CID, the POI was translocated from the
Golgi to the plasma membrane[12]. However, binding of the POI to
Golgi was difficult to control, as there was competition between
binding to the Golgi and to the plasma membrane. We sought to
solve this issue via a novel “parallel MAC” approach (Figure 2A,
Movie S1). Because the ST-induced dimerization is stable and
tunable (Figure S2, S6), ST could be used to tune the POI level
in the cytosol. After wash-out of ST, photo-activation of NvocTMP-
Cl induces dimerization. Since these two events are independent
to each other, we termed this approach “parallel MAC”.
To this end, three constructs were co-expressed in HeLa cells:
cytosolic Citrine-eDHFR-Rac1*, mCherry-HaloTag-CAAX at the
plasma membrane and mTurquoise2-2×FKBP'-NLS (nucleus
localization signal) in the nucleus (Figure 2A-B). Herein Rac1* (i.e.
Rac1Q61L∆CAAX) stands for a constitutively active Q61L mutant
that lacks plasma membrane localization.
Targeting of Rac1* to the plasma
membrane will activate WAVE, and the
Arp2/3 complex, which will stimulate actin
polymerization near the plasma membrane,
resulting in cell protrusion (Figure 2A). In
HeLa cells, those localized protrusions
mainly take the shape of dynamic ruffles at
the plasma membrane.[13] Citrine-eDHFR-
Rac1* can freely diffuse in the cytosol,
which can already cause significant
membrane ruffling prior to light-induced
dimerization (Figure 2B left panel, indicated
by arrows). In order to control the amount of
Rac1* in the cytosol, we added the first
dimerizer ST to recruit cytosolic Rac1* into
the nucleus. Titration of ST revealed a
dose-dependent
decrease
of
the
cytosol/nucleus ratio with a half maximal
effective concentration (EC50) of 0.53±0.04
μM, showing that cytosolic Rac1*
concentration can be fine-tuned by the
addition of ST (Figure S6). We found that
partial sequestration of Rac1* to the
nucleus by addition of 0.3 μM ST is
sufficient to minimize basal Rac1*-induced
ruffle formation (Figure 2B-E). Following
wash-out of ST, illumination at the edge of
the cell rapidly targeted Rac1* to the region
of photoactivation (ROP) at the plasma
membrane (Figure 2B, 2E) without
alteration of Rac1* in the nucleus (Figure
2D). This suggests that ST promotes stable
dimerization to sequester Rac1* in the
Figure 2. Parallel multi-directional activity control (“parallel MAC”) of Rac1 in live cells. A) Schematic
representation of the principle of the “parallel MAC” approach. B) Confocal images of a HeLa cell co-
expressing Citrine-eDHFR-Rac1* (cytosol), mCherry-HaloTag-CAAX (plasma membrane) and
mTurquoise2-2×FKBP'-NLS (nucleus). The rectangular area indicates the region of photoactivation
(ROP) by 405 nm light. Arrows indicate ruffles. Scale bar: 10 µm. C) Normalized fluorescence intensity
ratio of Citrine-eDHFR-Rac1* at the ROP versus the entire cell (n=4 cells). Ctirine-eDHFR-Rac1* level
in the cytosol decreased after adding 0.3 µM ST, and was then recruited to the ROP upon
photoactivation (PA). D) Normalized fluorescence intensity ratio of Citrine-eDHFR-Rac1* versus
mTurquoise2-2×FKBP'-NLS in the nucleus before (-ST) and after the addition of 0.3 μM ST for 20 min
(+ST), and after photoactivation (PA). n.s., not significant (p=0.46) from paired Student’s t-test. E) Time
series of images of the ROP shows the recruitment of Citrine-eDHFR-Rac1* (upper panel, intensity
coded) and ruffle formation (arrows) at the plasma membrane (lower panel, mCherry-HaloTag-CAAX).
For C) and D), mean values and standard error of the mean (SEM) were shown. See also Movie S1.
This article is protected by copyright. All rights reserved.

10.1002/anie.201806976
Angewandte Chemie International Edition
COMMUNICATION
nucleus, which is not reversible without the competitor (Figure S2). competitively inhibit the NvocTMP-Cl induced dimerization of
Light-induced plasma membrane targeting leads to significant
formation of ruffles (observed with the plasma membrane marker
mCherry-HaloTag-CAAX) only at the ROP (Figure 2B, right panel;
Figure 2E). Therefore, the “parallel MAC” approach using the
pdCID system is well suited to tune background activity before
light-induced control of protein function in cells.
eDHFR and HaloTag while trigger the S*T induced dimerization
of eDHFR and FKBP'. To enhance the efficiency of eDHFR/FKBP'
dimerization, we introduced the second-generation S*T dimerizer
that is optically pure (Figure 1 & Scheme S1). Compared to the
first generation ST (EC50 = 0.53±0.04 μM, Figure S6), which is a
diastereomeric mixture, S*T not only shows 8-fold higher efficacy
(EC50 = 0.065±0.005 µM), but also displays a higher extent of
protein dimerization in cells (Figure S7). First, we demonstrated
that S*T can disrupt the dimerization between eDHFR and
HaloTag that was initially induced by TMP-Cl (the uncaged form
of NvocTMP-Cl) with an EC50 of 3.2±0.2 µM (Figure S8),
suggesting that the “competitive MAC”
Another challenge in chemical genetic and optogenetic
systems is to achieve multiple layers of functional control in a
single cell. To address this problem, we developed the
“competitive MAC” approach to enable multi-layered control over
protein or organelle function. In this approach, S*T can
approach is feasible.
As a proof of principle, we used the
competitive MAC approach to cycle the
positioning of proteins among multiple sites
within a cell. Recent studies show that in
addition to the cytosol and plasma membrane,
Rac1 also localizes in the nucleus and
regulates nuclear membrane organization and
morphology. The nucleocytoplasmic shuttling
of Rac1 plays an important role in tumor
invasion.[14] Using the pdCID system, we
manipulated the shuttling of Rac1 among the
cytosol, plasma membrane and nucleus
(Figure S9, Movie S2). We also performed
multiple cycles of Rac1 shuttling among the
cytosol, mitochondria and nucleus (Figure
S10).
We further used the competitive MAC
approach to control bidirectional transport of
organelles by targeting two microtubule motors
with opposite directionality (Figure 3A). To this
end, three constructs, PEX3-mCherry-eDHFR
that anchors at the peroxisome through the
PEX3 peptide motif, KIF5BN-Citrine-HaloTag
that contains the N-terminal motor domain (1-
560) of the kinesin KIF5B (i.e. KIF5BN), and
BicD2N-mTurquoise2-2×FKBP’ that contains
the N-terminal dynein binding domain (1-594)
of the dynein adaptor protein Bicaudal D2 (i.e.
BicD2N), were co-expressed in HeLa cells.
Recruitment of kinesin (KIF5BN) or the dynein
Figure 3. Competitive multi-directional activity control (“competitive MAC”) of peroxisome transport.
A) Schematic representation of the principle of competitive MAC. B) Representative confocal
images. Cherry channel: the arrows indicate that peroxisomes cluster at the cell periphery; the
arrow head indicates that peroxisomes accumulate at the microtubule organization center (MTOC)
at the cell center. C) Pearson’s correlation coefficient (PCC) analysis of the colocalization between
KIF5BN-Citrine-eDHFR and PEX3-mCherry-HaloTag showed that KIF5BN is recruited to
peroxisomes after PA and then dissociates from peroxisomes after adding S*T. D) The
fluorescence intensity ratio of BicD2N at the MTOC versus the entire cell increases after adding
S*T and decreases after adding TMP. E) The fluorescence intensity ratio of PEX3 (peroxisome
marker) at the MTOC versus the entire cell increases after S*T, but does not change significantly
after adding TMP for 3 min. Scale bar, 25μm. For C), D), and E), mean values and SEM were
shown; data were based on n=6 cells from three independent experiments. p values are determined
by paired Student’s t-test (***: p < 0.001; **: p < 0.01; *: p < 0.05; n.s.: not significant); “Post PA”
indicates 5-10 min after PA and just before adding S*T, while “Post +S*T” means 5-10 min after the
addition of S*T and just before adding TMP. F) Time series of montages of an enlarged region
within the cell as shown in Figure S11. See also Movie S3 and S4.
complex (via BicD2N) to cargos induces
microtubule plus-end or minus-end directed
transport to the cell periphery or to the cell body,
respectively. NvocTMP-Cl was pre-bound to
KIF5BN via the HaloTag. Photoactivation (PA)
led to the recruitment of KIF5BN to
peroxisomes (Figure 3B, C), which induced
their anterograde transport to the cell periphery
(Figure 3B: Pre, PA 2', PA 5'). Subsequently,
addition of S*T led to the displacement of
KIF5BN and recruitment of BicD2N to
peroxisomes (Figure 3B: S*T 80", Figure 3C),
which stimulated retrograde transport of
This article is protected by copyright. All rights reserved.

10.1002/anie.201806976
Angewandte Chemie International Edition
COMMUNICATION
peroxisomes to the cell center (Figure 3B: S*T 9', Figure 3D-E,
Movie S3). This bidirectional peroxisome transport was readily
visualized within a subcellular region. Upon PA, dispersed
peroxisome vesicles moved to the cell periphery, where they
formed a cluster. Addition of S*T triggered the retrograde
transport of peroxisomes toward the cell body (Figure 3F, Figure
S11, Movie S4). Finally, TMP dissociated BicD2N from
peroxisomes (cyan channels of Figure 3B: TMP 80"  TMP 3',
Figure 3D) and disrupted the directional movement of
peroxisomes (red channels of Figure 3B: TMP 80"TMP 3',
Figure 3E). By swapping KIF5BN and BicD2N, BicD2N was first
recruited to cargos to induce retrograde motility, followed by
replacing BicD2N with KIF5BN to trigger anterograde transport
(Figure S12, Movie S5). Hence, the “competitive MAC” approach
can be flexibly implemented to control the direction of cargo
transport.
Acknowledgements
We thank Sven Müller for technical support in microscopy. This
work was supported by the Deutsche Forschungsgemeinschaft,
DFG (grant No.: SPP 1623), Behrens Weise Stiftung, European
Research Council, ERC (ChemBioAP) and Knut and Alice
Wallenberg Foundation to Y.W.W.
Keywords: chemo-optogenetics • MAC • pdCID • Rac1 •
bidirectional cargo transport
[1]
[2]
[3]
[4]
[5]
G. S. Butler, C. M. Overall, Nat. Rev. Drug Discov. 2009, 8, 935-948.
C. J. Jeffery, Mol. Biosyst. 2009, 5, 345-350.
B. T. MacDonald, K. Tamai, X. He, Dev. Cell 2009, 17, 9-26.
M. C. Hung, W. Link, J. Cell Sci. 2011, 124, 3381-3392.
a) P. van Bergeijk, C. C. Hoogenraad, L. C. Kapitein, Trends Cell Biol.
2016, 26, 121-134; b) P. van Bergeijk, M. Adrian, C. C. Hoogenraad, L.
C. Kapitein, Nature 2015, 518, 111-114.
Most cargos simultaneously bind to kinesin and cytoplasmic
dynein motors, which move in opposite directions along
microtubules. A model of a stochastic “tug-of-war” that describes
the mechanical competition between antagonistic motors bound
to the same cargo has been proposed.[15] Small changes in the
number of active motors lead to significant changes in the net
direction of cargo motility.[16] In our study, the extra force
generated by the recruitment of additional kinesin or dynein
motors outcompetes the force generated by the endogenous
motors, thereby enforcing cargo transport toward the cell
periphery or toward the cell center (Figure S13). Our “competitive
MAC” experiments provide direct experimental support for the
“tug-of-war” model, regardless how exactly dynein and kinesin
motors coordinate with each other at the molecular level.
[6]
[7]
a) W. O. Hancock, Nat. Rev. Mol. Cell Biol. 2014, 15, 615-628; b) K. J.
De Vos, A. J. Grierson, S. Ackerley, C. C. Miller, Annu. Rev. Neurosci.
2008, 31, 151-173.
a) A. Fegan, B. White, J. C. T. Carlson, C. R. Wagner, Chem. Rev. 2010,
110, 3315-3336; b) S. Voss, L. Klewer, Y. W. Wu, Curr. Opin. Chem. Biol.
2015, 28, 194-201; c) R. DeRose, T. Miyamoto, T. Inoue, Pflug. Arch.
Eur. J. Phy. 2013, 465, 409-417; d) M. Putyrski, C. Schultz, FEBS Lett.
2012, 586, 2097-2105.
[8]
[9]
a) D. Tischer, O. D. Weiner, Nat. Rev. Mol. Cell Biol. 2014, 15, 551-558;
b) M. Weitzman, K. M. Hahn, Curr. Opin. Cell Biol. 2014, 30, 112-120; c)
X. Chen, Y. W. Wu, Angew. Chem. Int. Ed. 2018, 57, 6796-6799; Angew.
Chem. 2018, 130, 6912-6915; d) N. Ankenbruck, T. Courtney, Y. Naro,
A. Deiters, Angew. Chem. Int. Ed. 2018, 57, 2768-2798; Angew. Chem.
2018, 130, 2816-2848.
a) R. Villasenor, Y. Kalaidzidis, M. Zerial, Curr. Opin. Cell Biol. 2016, 39,
53-60; b) M. Schmick, N. Vartak, B. Papke, M. Kovacevic, D. C. Truxius,
L. Rossmannek, P. I. H. Bastiaens, Cell 2014, 157, 459-471; c) C. Niehrs,
Nat. Rev. Mol. Cell Biol. 2012, 13, 767-779.
In this study, we have shown that the pdCID system can be
flexibly implemented in distinct perturbation strategies to achieve
multi-layered activity control in a single cell. Another advantage of
the pdCID system is its selectivity and full bioorthogonality.
NvocTMP-Cl and ST enable the combination of rapid, light-
induced (typically t1/2 < 1 s) and chemically induced (t1/2 < 1 min)
dimerization, respectively. Simultaneous combination of all these
desirable features has not been achieved in other CID systems.
[10] P. Liu, A. Calderon, G. Konstantinidis, J. Hou, S. Voss, X. Chen, F. Li, S.
Banerjee, J. E. Hoffmann, C. Theiss, L. Dehmelt, Y. W. Wu, Angew.
Chem. Int. Ed. 2014, 53, 10049-10055; Angew. Chem. 2014, 126, 10213
-10219.
[11] X. Chen, M. Venkatachalapathy, D. Kamps, S. Weigel, R. Kumar, M.
Orlich, R. Garrecht, M. Hirtz, C. M. Niemeyer, Y.-W. Wu, L. Dehmelt,
Angew. Chem. Int. Ed. 2017, 56, 5196-5920; Angew. Chem. 2017, 129,
6010-6014.
In summary, using the pdCID system, we developed a multi-
directional activity control (termed “MAC”) strategy. The “parallel
MAC” approach was used to address the inherent problem of
undesirable background activity in chemical/optogenetic systems
through fine-tuning of the cytosolic level of Rac1. The “competitive
MAC” approach enables reversible, and multiple layers of
functional control, which is exemplified by multi-directional control
of the positioning of organelles and proteins in a single cell. We
envision that these versatile MAC strategies open up new
avenues for spatiotemporal control of proteins or organelles,
which cannot be addressed with existing techniques. These new
tools are particularly useful to study how protein or organelle
function is modulated by their local, subcellular environment, and,
conversely, how proteins or organelles reciprocally affect their
local environment and eventually cellular function. The MAC
approach could also be used to emulate or interfere with disease
conditions involving protein/organelle positioning in order to study
pathogenic mechanisms, and ultimately aid the development of
their therapeutic intervention.
[12] S. C. Phua, C. Pohlmeyer, T. Inoue, ACS Chem. Biol. 2012, 7, 1950-
1955.
[13] Y. I. Wu, D. Frey, O. I. Lungu, A. Jaehrig, I. Schlichting, B. Kuhlman, K.
M. Hahn, Nature 2009, 461, 104-108.
[14] I. Navarro-Lerida, T. Pellinen, S. A. Sanchez, M. C. Guadamillas, Y. H.
Wang, T. Mirtti, E. Calvo, M. A. Del Pozo, Dev. Cell 2015, 32, 318-334.
[15] M. J. I. Müller, S. Klumpp, R. Lipowsky, P. Natl. Acad. Sci. USA 2008,
105, 4609-4614.
[16] A. G. Hendricks, E. Perlson, J. L. Ross, H. W. Schroeder, 3rd, M. Tokito,
E. L. Holzbaur, Curr. Biol. 2010, 20, 697-702.
This article is protected by copyright. All rights reserved.

10.1002/anie.201806976
Angewandte Chemie International Edition
COMMUNICATION
Entry for the Table of Contents (Please choose one layout)
Layout 1:
COMMUNICATION
Text for Table of Contents
Xi Chen, Muthukumaran
Venkatachalapathy, Leif Dehmelt, Yao-
Wen Wu*
Page No. – Page No.
Multi-directional activity control
(MAC) of cellular processes enabled
by a versatile chemo-optogenetic
approach
This article is protected by copyright. All rights reserved.