Signal transduction overview
Timothy T. Cornell, MD; Thomas P. Shanley, MD
P erhaps nothing is more critical
to the biology of life than the
ability of the single cell to
sense and respond to an exter-
nal stimulus. Changes in the extracellu-
lar environment or direct application of a
stimulus to the cell must be “sensed” at
the interface of the cell and its external
environment. This “stimulus” commonly
creates the need for the cell to mount
some kind of response, often necessitat-
ing initiation of the expression of a gene
through the process of transcription. In
order for the signal sensed at the cell
surface to be communicated to the tran-
scriptional machinery in the nucleus re-
quires the propagation or transduction of
this signal through the cytosol to the
nuclear compartment (Fig. 1). Although
cells have adapted a broad array of path-
ways to accomplish this biological task,
some general mechanistic principles are
conserved among all living organisms.
The term “signal transduction” defines
this function of those pathways that have
evolved to accomplish this fundamental
cellular process. Other terms that have
been used to describe this biological
function include “cell signaling” or
“transmembrane signaling.”
Although this field now comprises its
own scientific entity, the science of signal
transduction owes its origin to the field of
endocrinology and hormonal biology.
The concept of a mechanism for intracel-
lular communication or “messaging”
originated from Claude Bernard’s work in
the 1850s, which described how “internal
secretions” of the ductless glands of the
thyroid and adrenals were released into
the circulation with consequent effects
on target organs. At the turn of the cen-
tury, this concept was refined by Bayliss
From the Division of Pediatric Critical Care Medi-
cine, C. S. Mott Children’s Hospital, University of Mich-
igan, Ann Arbor, MI.
Copyright © 2005 by the Society of Critical Care
Medicine and Lippincott Williams & Wilkins
DOI: 10.1097/01.CCM.0000191713.71308.FD
S410
and Starling (1) in their initial descrip-
tion of “secretin,” which was identified as
a member of a larger group of chemical
messengers that came to be called “hor-
mones,” a term purportedly coined by
W. B. Hardy in 1905 (2). This was the first
work to summarize the biological phe-
nomenon of a secreted product affecting
a characteristic response in target cells,
marking this year (2005) as the 100th
anniversary of our initial understanding
of signal transduction. During the most
recent decades, major advances in molec-
ular biology and imaging have vastly in-
creased our understanding of signal
transduction pathways at every level,
ranging from those mechanisms utilized
by various receptor types that recognize a
stimulus, through the array of second
messengers that propagate the signal,
culminating in the numerous nuclear
factors that alter gene expression. It is
the goal of subsequent articles to focus
on many of these specific pathways,
whereas the intention of this article is to
familiarize the reader with the general
mechanisms of signal transduction, par-
ticularly as they pertain to inflammatory
cell signaling and those disease states ad-
dressed on a daily basis by practicing in-
tensivists.
As suggested above, the process of sig-
nal transduction involves a series of con-
served mechanisms arranged in a path-
way framework that must transmit a
signal to the machinery that creates a
cellular response typically resulting in an
alteration of gene expression within the
nuclear compartment (Fig. 2). The first
step in this process involves the initiation
of a signal in response to an extracellular
or membrane event that generally re-
quires a receptor or cell membrane
change to sense this stimulus. Next, a
series of intracellular protein-to-protein
signal propagating events occur to trans-
mit the signal through the cytosol to the
nucleus. Numerous effectors are em-
ployed for the purpose of signal propaga-
tion, including the reversible phosphory-
lation of proteins, the mobilization of
sequestered ionized calcium, the activa-
tion of lipid-derived mediators, the accu-
mulation or degradation of cyclic nucle-
otides (namely, cyclic adenosine
monophosphate or cyclic guanosine
monophosphate), and stimulation of G-
coupled proteins. Finally, the principal
function of these pathways is to allow for
regulation of de novo gene expression
effecting the production of proteins that
ultimately characterize the cellular re-
sponse. This latter process starts in the
nuclear compartment via activation of
the transcriptional machinery (discussed
in the accompanying article, “Transcrip-
tion”). In light of the myriad of receptors,
adaptor proteins, kinases, phosphatases,
and other components that comprise this
vast number of pathways, a full under-
standing of the complexity of even a sin-
gle pathway can be a daunting challenge.
Thus, in providing the reader with brief
examples of each of these steps in signal
transduction, we have highlighted those
pathways with specific relevance to the
biology of critical illness.
Signal transduction is initiated by any
number of stimuli that influence cellular
responses and activities. Among those
stimuli relevant to critical care are circu-
lating mediators (hormones, cytokines,
growth factors), osmolar changes, me-
chanical stress (such as shear, stretch)
(3), and pathogens. Of particular interest
to intensivists is the response triggered
by the presence of a bacterial pathogen
that mediates the pathophysiologic clini-
cal responses of sepsis. Over the past de-
cade, investigators have identified char-
acteristic patterns expressed by
pathogens that make them unique from
the human host. These patterns have
been called pathogen-associated molecu-
lar patterns or PAMPs. To sense these
unique patterns, hosts have evolved a
mechanism to recognize them. The Toll-
like receptors, which have specific affinity
for the various pathogen products, are
fundamental to this process (4). Toll-like
Crit Care Med 2005 Vol. 33, No. 12 (Suppl.)
Figure 1. Principles of signal transduction. mRNA, messenger ribonucleic acid.
receptors are similar to most receptors in
that they are embedded in the cell mem-
brane and possess an extracellular ligand-
binding domain and usually an intracel-
lular signaling domain (5). Alternatively,
some receptors may utilize a second pro-
tein possessing an extension into the cell
to form a “receptor complex,” as exem-
plified by the Toll-like receptor 4/lipo-
polysaccharide receptor. Using this
mechanism, Toll-like receptors are able
to transmit the signal sensed at the cell
surface into the internal milieu of the
cell.
Once the signal has been transmitted
from the extracellular space through the
cell membrane, any number of serial or
parallel transducing proteins or amplifi-
ers can serve as additional points for reg-
ulating the propagation of the signal.
Perhaps the most common mechanism
by which proteins propagate a signal in
mammalian cells is by reversible phos-
phorylation of serine, threonine, or ty-
rosine residues (6). Protein kinases,
which form one of the largest gene fam-
ilies (⬎1000 genes), catalyze the transfer
of a ␥-phosphate group from adenosine
triphosphate to these specific amino acid
residues. Relevant examples of serine/
Crit Care Med 2005 Vol. 33, No. 12 (Suppl.)
threonine kinases that modulate inflam-
matory responses associated with sepsis
and acute lung injury are the mitogen-
activated protein kinases that include the
extracellular signal–regulated kinase, c-
Jun N-terminal kinase, and p38 pathways
(7, 8). Protein tyrosine kinases are di-
vided into two main classes: receptor and
cellular or nonreceptor protein tyrosine
kinases. With receptor protein tyrosine
kinases, such as the fibroblast growth fac-
tor receptor, ligand binding at the extra-
cellular domain causes receptor dimer-
ization and autophosphorylation of
intracellular tyrosine residues, which
provide docking sites for additional
adapter signaling proteins. In contrast,
cellular or nonreceptor protein tyrosine
kinases are cytosolic (or even nuclear) in
location and are activated similarly to the
classic serine/threonine kinases. An ex-
ample of this group of kinases is interleu-
kin-10 –mediated activation of a janus-
activated kinase that phosphorylates
STAT proteins to mediate an anti-
inflammatory effect (9).
Experimentally, activation of a specific
kinase is generally measured in two ways,
either by examining cellular proteins for
evidence of phosphorylation of the target
substrate using Western blot analysis or
by direct measurement of the kinase ac-
tivity. The latter is performed by immu-
noprecipitating the enzyme of interest
followed by an in vitro kinase assay in
which the kinase’s substrate and a radio-
labeled source of phosphate (phospho-
rus-32 adenosine triphosphate) are added
such that phosphorylation of the sub-
strate can be determined by radiography.
Although both methods are commonly
employed, it is important to emphasize a
caveat when relying on Western blot
analysis alone for estimating kinase activ-
ity. It must be recognized that at any
point in time, the phosphorylation state
of a protein is balanced between the phos-
phorylation by kinases and the dephos-
phorylation by a family of enzymes called
phosphatases capable of dephosphorylat-
ing it. The phosphates are composed of a
large family of proteins that hydrolyze
the phosphoester bonds of phosphory-
lated serine, threonine, and tyrosine res-
idues and thus are essential components
of the process of reversible protein phos-
phorylation. Two large protein phospha-
tase families exist: serine/threonine and
tyrosine phosphatases. Examples from
each family include the Ser/Thr protein
S411
Figure 2. General mechanisms employed by signal transduction pathways. TF, transcription factor.
phosphatase, PP2A, that regulates c-Jun
N-terminal kinase activity (10) and the
tyrosine protein phosphatase, mitogen-
activated protein kinase phosphatase-1,
that regulates p38 activity and mediates
endotoxin tolerance (11). As a result, con-
firmation of specific kinase activity
should be attempted in all cases to cor-
roborate increases or decreases in the
phospho-state of a protein determined by
Western blot analysis.
As mentioned above, additional means
of signal propagation include calcium
mobilization, activation of lipid-derived
mediators, changes in cyclic nucleotide
(e.g., cyclic adenosine monophosphate)
concentration, and stimulation of G-
coupled proteins. In some manner, all of
these mechanisms are relevant to critical
care. Myocyte activation by depolariza-
tion signals the release of calcium from
the sarcoplasmic reticulum to facilitate
S412
contraction of the myocyte. Recent data
show that platelet activating factor in-
creases the sphingolipid product, cer-
amide, resulting in activation of the nu-
clear factor-␬B and AP-1 pathways,
resulting in experimental pulmonary
edema (12). The potent vasoconstrictor
effect of endothelin-1 observed with
pathologic pulmonary hypertension is
mediated by the G-protein– coupled, ETA
endothelin receptor (13). Numerous ad-
ditional examples throughout the critical
care literature examine pathways utiliz-
ing these common mechanisms, al-
though it is beyond the scope of this
article to describe them in detail.
Although much of the focus on signal
transduction studies has been aimed at
elucidating the “normal” functions of the
cell, our understanding of the role of sig-
nal transduction pathways in mediating
pathologic or detrimental responses (e.g.,
cancer cell transformation, dysregulated
inflammation) has substantially in-
creased during the past decade. To this
end, some pathways have been examined
in preliminary studies of sepsis and acute
lung injury to determine their contribu-
tion to the observed clinical pathophysi-
ologic states. As an example, a working
hypothesis suggests that genes regulated
by the nuclear factor-␬B pathway (see
accompanying article) mediate multiple
organ dysfunction syndrome in critically
ill patients. In a recent clinical study,
baseline levels of cellular nuclear fac-
tor-␬B were higher in the nonsurvivors of
sepsis as compared with the survivors. In
addition, the derived receiver operating
characteristic curve showed cellular nu-
clear factor-␬B signal to be a better pre-
dictor of mortality than the Acute Physi-
ology and Chronic Health Evaluation II
score (14). There is little doubt that fu-
Crit Care Med 2005 Vol. 33, No. 12 (Suppl.)
ture advances in our understanding of
those pathways involved in the patho-
physiology of diseases such as sepsis and
acute lung injury will afford better strat-
ification of patients for clinical trials and
additional targets for novel therapeutic
approaches.
Conclusion
Signal transduction pathways com-
prise the means by which the cell senses
a stimulus and transmits that message to
other machinery capable of instituting a
cellular response. The complexity and in-
terconnectedness of these pathways cre-
ate a challenge for investigators and read-
ers alike in deciphering their precise
roles. Many observations are not only cell
specific but also stimulus or model spe-
cific. With such a complex series of path-
ways regulating innumerable cellular
functions as elucidated with single-cell
models, how can we determine the in
vivo role of signaling in affecting our
patients with critical illnesses? Although
this remains a daunting challenge, our
goal of identifying novel therapeutic ap-
proaches in sepsis and acute respiratory
distress syndrome is dependent on this
translational effort. As we proceed, we
must recognize certain experimental lim-
itations. Because both kinases and phos-
phatases regulate reversible phosphoryla-
tion, it behooves researchers to consider
Crit Care Med 2005 Vol. 33, No. 12 (Suppl.)
the effect of activation or inhibition of
these enzymes on the level of phosphor-
ylated proteins observed. Thus, direct
measurement of kinase/phosphatase ac-
tivity should be the primary goal. Finally,
signal propagation by a specific pathway
should result in a downstream cellular
response (e.g., gene expression, growth
or differentiation, motility). In drawing
conclusions from studies elucidating
these pathways, authors should be re-
porting a downstream “readout” result-
ing from either activation or inhibition of
the pathway. As our methodologic ap-
proaches evolve and improve, especially
with proteomic approaches affording the
examination of posttranslational protein
changes, our understanding of signaling
pathways can be anticipated to continue
to grow at a tremendous pace. However,
as signaling is the language that the cell
speaks, the task facing the critical care
community is to one day become fluent
in this language.
REFERENCES
1. Bayliss WM, Starling EH: The mechanism of
pancreatic secretion. J Physiol 1902; 28:325
2. Wright RD: The origin of the term “hor-
mone.” Trends in Biochem Sci 1978; 3:275
3. dos Santos CC, Slutsky AS: Mechanotrans-
duction, ventilator-induced lung injury and
multiple organ dysfunction syndrome. Inten-
sive Care Med 2000; 26:638 – 642
4. Means TK, Golenbock DT, Fenton MJ: Struc-
ture and function of Toll-like receptor pro-
teins. Life Sci 2000; 68:241–258
5. Dunne A, O’Neill LA: The interleukin-1 re-
ceptor/Toll-like receptor superfamily: Signal
transduction during inflammation and host
defense. Sci STKE 2003;171:re3
6. Hunter T: Protein kinases and phosphatases:
The yin and yang of protein phosphorylation
and signaling. Cell 1995; 80:225–236
7. Chang L, Karin M: Mammalian MAP kinase
signaling cascades. Nature 2001; 410:37– 40
8. Johnson GL, Lapadat R: Mitogen-activated
protein kinase pathways mediated by ERK,
JNK, and p38 protein kinases. Science 2002;
298:1911–1912
9. Donnelly RP, Dickensheets H, Finbloom DS:
The interleukin-10 signal transduction path-
way and regulation of gene expression in
mononuclear phagocytes. J Interferon Cyto-
kine Res 1999; 19:563–573
10. Shanley TP, Vasi N, Denenberg AG, et al: The
serine/threonine phosphatase, PP2A: Endog-
enous regulator of inflammatory cell signal-
ing. J Immunol 2001; 166:966
11. Nimah M, Zhao B, Denenberg AG, et al: Con-
tribution of MKP-1 regulation of p38 to en-
dotoxin tolerance. Shock 2005; 23:80 – 87
12. Goggel R, Winoto-Morbach S, Vielhaber G, et
al: PAF-mediated pulmonary edema: A new
role for acid sphingomyelinase and ceramide.
Nat Med 2004; 10:155–160
13. Galie N, Manes A, Branzi A: The endothelin
system in pulmonary arterial hypertension.
Cardiovasc Res 2004; 61:227–237
14. Arnalich F, Garcia-Palomero E, Lopez J, et al:
Predictive value of nuclear factor kappaB ac-
tivity and plasma cytokine levels in patients
with sepsis. Infect Immun 2000; 68:
1942–1945
S413