Perrimon, Norbert

BMC Genomics

Norbert Perrimon

Springer - Publisher Connector

Signaling in time and space

ORAL PRESENTATION Open Access
Signaling in time and space
Norbert Perrimon
From 2nd International Genomic Medical Conference (IGMC 2013)
Jeddah, Kingdom of Saudi Arabia. 24-27 November 2013
Organisms sense and interpret numerous internal and
external signals by activating various signal transduction
pathways. Past studies have illustrated that not only the
presence or absence of one signal matters, but also that
the frequency, amplitude, and duration of the signal pro-
vide crucial information to control many biological pro-
cesses such as DNA repair, cell cycle, cell proliferation and
differentiation. For example, oscillation of p35 activity is
associated with cell-cycle arrest and recovery, whereas a
single long-lasting activation of p35 leads to apoptosis. As
a result, it is critical to have methods for detection of sig-
nal activity in vivo and in real time. Although the applica-
tion of genetically encoded fluorescent proteins (FPs) as
live signal reporters has revolutionized the field, most of
the FP-based transcriptional reporters suffer from slow
maturation (>1 hr) and extremely long half-life (>24 hr).
This limitation greatly reduces the ability of these repor-
ters to reveal the real-time dynamics of signals. To address
this issue, new FP variants such as superfolder GFP
(sfGFP) with a faster maturation time and high brightness
have been engineered. In addition, the half-life of FPs can
be shortened to about 2 hr by tagging them with a PEST
domain for protein degradation. However, although short-
lived FPs can report changes more faithfully, they have a
major drawback in that they show a dramatic decrease
in the fluorescent signal since much less FP is left for
detection. I will discuss our recent efforts at developing
new FP with improved temporal resolution.
In addition, our laboratory is exploring methods to
interrogate the organization of protein complexes in
polarized cells. Polarization of the epithelial cell defines
its functions, including selective absorption, secretion,
and transport. Along the apical to basal axis, at least five
distinct membrane domains can be defined. Numerous
methods have been developed to determine the protein
composition of polarized plasma membrane domains in
epithelia. However, results to date have been inadequate,
and have yielded relatively few insights into the biogen-
esis or functional differentiation of these domains. I will
discuss our recent efforts at implementing in vivo meth-
ods of live cell proteomics using engineered peroxidase.
Published: 2 April 2014
doi:10.1186/1471-2164-15-S2-O18
Cite this article as: Perrimon: Signaling in time and space. BMC
Genomics 2014 15(Suppl 2):O18.
Submit your next manuscript to BioMed Central
and take full advantage of: 
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at 
www.biomedcentral.com/submitCorrespondence: perrimon@rascal.med.harvard.edu
Department of Genetics, Harvard Medical School, USA
Perrimon BMC Genomics 2014, 15(Suppl 2):O18
http://www.biomedcentral.com/1471-2164/15/S2/O18
© 2014 Perrimon; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited.


Harvard University - DASH 

 Signaling in time and space  (Article begins on next page)The Harvard community has made this article openly available.Please share how this access benefits you. Your story matters.Citation Perrimon, Norbert. 2014. “Signaling in time and space.” BMCGenomics 15 (Suppl 2): O18. doi:10.1186/1471-2164-15-S2-O18. http://dx.doi.org/10.1186/1471-2164-15-S2-O18.Published Version doi:10.1186/1471-2164-15-S2-O18Accessed February 16, 2015 4:46:17 PM ESTCitable Link http://nrs.harvard.edu/urn-3:HUL.InstRepos:12717597Terms of Use This article was downloaded from Harvard University's DASHrepository, and is made available under the terms and conditionsapplicable to Other Posted Material, as set forth athttp://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAAORAL PRESENTATION Open AccessSignaling in time and spaceNorbert PerrimonFrom 2nd International Genomic Medical Conference (IGMC 2013)Jeddah, Kingdom of Saudi Arabia. 24-27 November 2013Organisms sense and interpret numerous internal andexternal signals by activating various signal transductionpathways. Past studies have illustrated that not only thepresence or absence of one signal matters, but also thatthe frequency, amplitude, and duration of the signal pro-vide crucial information to control many biological pro-cesses such as DNA repair, cell cycle, cell proliferation anddifferentiation. For example, oscillation of p35 activity isassociated with cell-cycle arrest and recovery, whereas asingle long-lasting activation of p35 leads to apoptosis. Asa result, it is critical to have methods for detection of sig-nal activity in vivo and in real time. Although the applica-tion of genetically encoded fluorescent proteins (FPs) aslive signal reporters has revolutionized the field, most ofthe FP-based transcriptional reporters suffer from slowmaturation (>1 hr) and extremely long half-life (>24 hr).This limitation greatly reduces the ability of these repor-ters to reveal the real-time dynamics of signals. To addressthis issue, new FP variants such as superfolder GFP(sfGFP) with a faster maturation time and high brightnesshave been engineered. In addition, the half-life of FPs canbe shortened to about 2 hr by tagging them with a PESTdomain for protein degradation. However, although short-lived FPs can report changes more faithfully, they have amajor drawback in that they show a dramatic decreasein the fluorescent signal since much less FP is left fordetection. I will discuss our recent efforts at developingnew FP with improved temporal resolution.In addition, our laboratory is exploring methods tointerrogate the organization of protein complexes inpolarized cells. Polarization of the epithelial cell definesits functions, including selective absorption, secretion,and transport. Along the apical to basal axis, at least fivedistinct membrane domains can be defined. Numerousmethods have been developed to determine the proteincomposition of polarized plasma membrane domains inepithelia. However, results to date have been inadequate,and have yielded relatively few insights into the biogen-esis or functional differentiation of these domains. I willdiscuss our recent efforts at implementing in vivo meth-ods of live cell proteomics using engineered peroxidase.Published: 2 April 2014doi:10.1186/1471-2164-15-S2-O18Cite this article as: Perrimon: Signaling in time and space. BMCGenomics 2014 15(Suppl 2):O18.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color ﬁgure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionSubmit your manuscript at www.biomedcentral.com/submit Correspondence: perrimon@rascal.med.harvard.eduDepartment of Genetics, Harvard Medical School, USAPerrimon BMC Genomics 2014, 15(Suppl 2):O18http://www.biomedcentral.com/1471-2164/15/S2/O18© 2014 Perrimon; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited.

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