The Evolution of Eukaryotic Cilia

and Flagella as Motile and Sensory

Organelles
David R. Mitchell*

Abstract

E
ukaryotic cilia and flagella are motile organelles built on a scaffold of doublet
microtubules and powered by dynein ATPase motors. Some thirty years ago, two
competing views were presented to explain how the complex machinery of these motile
organelles had evolved. Overwhelming evidence now refutes the hypothesis that they are the
modified remnants of symbiotic spirochaete-like prokaryotes, and supports the hypothesis that
they arose from a simpler cytoplasmic microtubule-based intracellular transport system.
However, because intermediate stages in flagellar evolution have not been found in living
eukaryotes, a clear understanding of their early evolution has been elusive. Recent progress in
understanding phylogenetic relationships among present day eukaryotes and in sequence analysis
of flagellar proteins have begun to provide a clearer picture of the origins of doublet and triplet
microtubules, flagellar dynein motors, and the 9+2 microtubule architecture common to these
organelles. We summarize evidence that the last common ancestor of all eukaryotic organisms
possessed a 9+2 flagellum that was used for gliding motility along surfaces, beating motility to
generate fluid flow, and localized distribution of sensory receptors, and trace possible earlier
stages in the evolution of these characteristics.

Evidence for the Presence of a 9+2, Motile, Sensory Organelle

in the Last Common Eukaryotic Ancestor
As summarized in Figure 1, typical cilia and flagella (hereafter called flagella, there being no
consistent structural or functional difference between organelles with these two designations)
are motile projections oriented perpendicular to the cell surface, but they vary in length, in
number per cell, and in the patterns of motility that they produce. They are composed of a
cylinder (the axoneme) of nine doublet microtubules surrounding two single microtubules and
are covered by the cell membrane. Between each pair of flagellar doublets are rows of axonemal
dynein ATPases, which power the bending of these organelles, and extending toward the center
of the cylinder are radial spokes, which touch upon a central apparatus and regulate axonemal
dyneins. This central element consists of two single microtubules, assembled from a unique
nucleating site,1 plus many interconnecting microtubule-associated proteins (reviewed in ref.

*Corresponding Author: David R. Mitchell—Department of Cell and Developmental

Biology, SUNY Upstate Medical University, 750 E. Adams St., Syracuse, New York
13210, U.S.A. Email: [email protected]

Origins and Evolution of Eukaryotic Endomembranes and Cytoskeleton,

edited by Gáspár Jékely. ©2006 Eurekah.com.
2 Origins and Evolution of Eukaryotic Endomembranes and Cytoskeleton

Figure 1. Diagram of structures common to all motile cilia and flagella. Longitudinal view to the
left shows the relationship between the axoneme and basal body, and the location of intraflagellar
transport (IFT) motors between axonemal doublet microtubules and the flagellar membrane.
Transition fibers attached to the basal body separate the flagellar membrane domain from the rest
of the cell membrane. Cross sectional views to the right show structures in flagella, including the
nine outer doublet and two single central pair microtubules (top) and the nine triplet microtu-
bules of basal bodies (bottom).

2). Together they form a structure that provides a cylindrical surface apposing the ends of the
radial spokes.3 The nine doublets assemble from a much shorter cylinder of nine triplet micro-
tubules, the basal body or centriole, which is anchored to the cell surface and stabilized in the
cytoplasm by other cytoskeletal elements. Basal bodies that anchor flagella are often inter-
changeable during the cell cycle with centrioles,4 and these two names should be considered as
two functional descriptions for the same structure. Between the doublets and the membrane
are particles associated with intraflagellar transport (IFT), a process important for flagellar
assembly and protein trafficking in this cellular compartment.5,6 In the outward (anterograde)
direction, IFT is powered by kinesins of the kinesin2 family; in the inward (retrograde) direc-
tion, power is provided by dyneins of the cytoplasmic dynein 2 (DHC1b) family. Many flagella
act as sensory antennae through localization of receptors to the flagellar membrane. In extreme
cases, termed primary cilia or sensory cilia, the motile function has been discarded and with it
the dyneins, radial spokes, the central pair complex, and other proteins needed for bend forma-
tion. IFT is still required for the assembly and maintenance of these primary cilia, which play
important sensory roles in metazoan organisms.6
Some attempts have been made in the past to identify intermediates in the evolution of
flagella by looking within existing branches of eukaryotes for organisms that may have diverged
before the complete 9+2 flagellum had evolved. However, improved methods of analysis and
the recent burst of sequence data are rapidly transforming long-held views of eukaryotic
phylogeny to new schemes in which there are many branches that diverged within a relatively
short period of time7,8 (Fig. 2). Many of these branches are represented today by single-celled
protists, so that the true diversity of eukaryotes cannot be appreciated without some under-
standing of the relationships among these often less-studied organisms. Thus the nearest
The Evolution of Eukaryotic Cilia and Flagella as Motile and Sensory Organelles 3

Figure 2. Diagram of probable evolutionary divergence that generated all existing branches of
eukaryotic organisms. Under the name of each branch or clade is a the name of a representative
genus in that clade that contains species with typical motile 9+2 flagella. Based on recent studies
of rare gene fusion events, as well as more traditional sequence comparisons, the entire tree is
divided into two superclades, unikonts and bikonts.

relatives of animals (metazoans) are single-celled choanoflagellates.9,10 Fungi (some of which

were once considered primitive because of their simplicity) turn out to be another twig of this
same branch, the opisthokonts or unikonts. Allomyces, a chytridiomycete fungus with
flagellated gametes and zoospores, is just one example of a fungal cell that swims, like a sperm,
with a typical 9+2 flagellum. Amoebozoa, containing such amoeboflagellates as Physarum, are
probably (based on a shared gene fusion and on mitochondrial sequences) the only other unikont
twig, and branch somewhat earlier than fungi.10-13 The other major superclade of eukaryotes,
the bikonts, encompasses a great variety of flagellated and amoeboid organisms, including
green plants and green and red algae (plantae), ciliates, dinoflagellates and their kin
(chromalveolata), euglenids, trypanosomatids, diplomonads and their sister taxa (excavata),
and the radiolaria, cercozoa, etc. (rhizaria).14,15 In the resulting tree (Fig. 2), one should note
that most of the model organisms under intense study during the past twenty years reside on
one branch (unikont), but fortunately for studies of flagellar evolution, additional attention
has been focused on a few flagellated bikont organisms, most especially the green alga
Chlamydomonas reinhardtii.
One striking conclusion of these recent phylogenetic studies is that every extant branch of
eukaryotes includes organisms with motile, 9+2 flagella. Even proteins of the central pair
apparatus, such as products of the Chlamydomonas PF6, PF16, PF20, KLP1, and CPC1 genes,
have been conserved between algae and humans.16 From this we can only conclude that these
organelles had evolved prior to the divergence of all extant eukaryotic clades from a common
ancestor. In addition, IFT proteins, which are central to flagellar assembly and to the display of
sensory receptors and flagellar surface motility, are also present in flagella from distant branches
of eukaryotic phylogeny (e.g., trypanosomes,17 insects,18 and green algae19), and therefore must
have evolved prior to the beginnings of eukaryotic radiation.18 The microtubule rootlet
structures that stabilize basal bodies in the cytoplasm do vary phylogenetically and therefore
the nature of those that might have been present in the last common eukaryotic ancestor are
difficult to determine,20,21 but all of the elements of triplet microtubules, and the accessory
proteins needed for basal body formation, must also have been present at the base of this tree.
Eukaryotes likely developed the nucleus, endomembrane system, and cytoskeleton, and
then used the phagocytic ability that was provided by the combined cytoskeletal and
4 Origins and Evolution of Eukaryotic Endomembranes and Cytoskeleton

endomembrane systems to obtain the precursors to mitochondria, long before the evolution of
flagella. The framework of doublet microtubules upon which flagella depend must have evolved
from simpler single microtubules which, as essential elements of the mitotic machinery, would
have been needed to segregate a genome enclosed in a nucleus. Likewise, dyneins as
microtubule-based motors undoubtedly functioned as transport motors on cytoplasmic and
mitotic microtubules long before their use was adapted to flagella. If all of the essential
elements of eukaryotic cells were in place for so long before the advent of flagella, one must ask
why there are no branches of existing eukaryotes that lack flagella. The simplest explanation is
that the branch of early eukaryotes that first developed a functional 9+2 flagellum possessed a
tremendous selective advantage over its competitors, and was the only eukaryote whose
descendants survive today.

Evolution of Tubulin, Dynein and Kinesin

The closest prokaryotic relative of tubulins, FtsZ, functions during bacterial septation, and
FtsZ homologs continue to perform a similar role in chloroplasts and perhaps some mitochon-
dria.22-24 In early eukaryotes, FtsZ gene duplication and modification led to alpha and beta
tubulin, which form a stable dimer that retains FtsZ properties such as polymerization,
GTP-binding, and GTP hydrolysis-dependent conformational change, but which gained the
added ability to interact laterally to form tubes. Gamma tubulin is also ubiquitous and likely
emerged early, to function as a nucleating site that helps determine microtubule polarity and
distribution. Additional tubulin isoforms delta and epsilon are ubiquitous among organisms
with triplet microtubules, and the formation of triplet microtubules, essential for the function
of basal bodies, has been shown to require both delta and epsilon tubulin in Chlamydomo-
nas25,26 and both epsilon27 and the less ubiquitous eta tubulin28 in Paramecium. The presence
of these tubulin isoforms in members of both the unikont and bikont clades17,29 argues for
their evolution prior to the divergence of eukaryotes.
The dynein motors that power both flagellar beating and retrograde IFT movements are
members of the superfamily of AAA ATPases. In dyneins, six individual AAA domains have
become fused into a single large polypeptide,30 but only four of these six domains in dyneins
retain the signature sequences of nucleotide binding pockets.31 DNA pumping ATPases of
archaea (HerA) and bacteria (FtsK), also AAA ATPases, are needed during prokaryotic cell
division for correct daughter chromosome segregation,32 and it would be tempting to assume
that an interaction between FtsK and FtsZ could have evolved directly into an interaction
between dynein and tubulin. However, dyneins apparently evolved from an entirely different
branch of the AAA superfamily from the DNA pumping ATPases. The closest eukaryotic rela-
tives of dyneins are midasins, similarly giant eukaryotic AAA ATPases, which function at the
nuclear pore in 60S ribosome export. The closest prokaryotic homologs of dyneins and midasins
are members of the MoxR family, single AAA domain ATPases that function as chaperones in
the assembly of large protein complexes, such as methanol dehydrogenase and nitric oxide
reductase.30 It would thus appear that dyneins evolved as microtubule motors in the early
eukaryotic lineage, and that their prokaryotic ancestors were proteins that performed confor-
mational work linked to ATP binding and hydrolysis.
Sequence comparisons among extant dynein heavy chains divide dyneins into two broad
families, cytoplasmic and axonemal.33 Most organisms have only two cytoplasmic dyneins, one
devoted to general cytoplasmic microtubule-based movements, present in all eukaryotes, and
one for retrograde IFT movement on axonemal microtubules, absent from organisms such as
Saccharomyces cerevisiae that lack axonemes. Each is thought to function as a homodimer of
catalytic heavy chains. Organisms with motile flagella also have a large family of axonemal
dyneins that can be further divided into three subfamilies, outer row dyneins, I1 inner row
dyneins, and additional diverse inner row dyneins (reviewed in ref. 34). Outer row dyneins
diverged before the common ancestor into two isoforms (alpha and beta) that form heavy
chain heterodimers. A third isoform that diverged more recently is found in Chlamydomonas
The Evolution of Eukaryotic Cilia and Flagella as Motile and Sensory Organelles 5

(plantae) and Tetrahymena (chromalveolata) but not in sea urchins or fruit flies (animalia).
Outer row dyneins bind in a continuous row with 24 nm spacing (every third 8 nm tubulin
dimer), whereas inner row dynein isoforms occur once every 96 nm along each doublet micro-
tubule.35 The I1 inner row dynein is a typical heterodimer of two heavy chain subunits, and
both isoform subfamilies are represented in the genomes of all organisms that retain motile
flagella. I1 dyneins appear to have become established early as major targets of signal-dependent
regulation of flagellar bending parameters.36 The many additional inner row dyneins present
in, for example, sea urchins and ciliates, appear to have diverged more recently; several of the
ciliate inner row dyneins are more closely related to each other than to any of the urchin inner
row dyneins. Structural, genetic and biochemical analyses in Chlamydomonas indicate that
these additional inner row dyneins function as monomers, rather than the dimers typical of all
other dyneins, and some isoforms may be differentially distributed along the length of the
organelle.37 Their sequence relationships suggest that the last common eukaryote may have
had a single isoform of this monomeric inner row dynein.
Among the members of the very large and diverse superfamily of kinesin ATPases are at
least two families with members that function as flagellar proteins. The small kinesin9 family is
represented by a sequence expressed in ciliated cells of mammals38 and by the Chlamydomonas
Klp1 protein.39 Klp1 has been localized to the central pair complex39 and shown to be impor-
tant for normal flagellar motility in that organism.40 Although evidence for the role of kinesin9
members in mammalian cells is not available, their expression patterns support an early evolv-
ing flagellar function for this protein. The kinesin2 family is larger and functionally more
diverse. While some members of the kinesin2 family are anterograde motors for IFT, others are
anterograde motors in other cytoplasmic compartments such as neurons of metazoans. The
presence of kinesin2 homologs in ciliates and flagellates, but not in nonflagellated fungi,
suggests that kinesin2 coevolved with axonemes and was only coopted for other transport
functions in recent metazoan evolution.6

The Origins of 9+2 Flagella

Given the evidence summarized above, the last common eukaryotic ancestor had a motile
9+2 flagellum, was anchored on a basal body of triplet microtubules, and required IFT for its
assembly. How did such a complex system evolve? Clearly it must have been preceded by a
microtubule cytoskeleton with dynein and kinesin motors. Strong arguments have been made
that the driving force for the evolution of a microtubule cytoskeleton and its associated motor
proteins was the ability to accurately segregate a large, nuclear membrane-enclosed genome by
mitosis. Although the complicated checks and balances used to assure mitotic fidelity vary
widely in extant eukaryotes, some aspects of mitosis have been sufficiently conserved and are so
central to the process that they must have been present at an early stage. Other aspects of
mitosis, assumed to be ancient because of their simplicity, reveal a minimal apparatus but may
not reflect the ancestral condition. Although model organisms such as fungi have provided
many details of such minimal systems, the tremendous variety of extant mitotic mechanisms
(as reviewed in ref. 41) should not be forgotten. Mitosis requires a microtubule organizing
center (MTOC) that duplicates once per cell cycle, a connection between each chromatid and
one of the duplicated MTOCs, and separation of the MTOCs with their associated chromatids.
These MTOCs vary structurally from the simple nuclear membrane-embedded spindle pole
bodies of some unicellular organisms to the complex centriole-containing centrosomes of many
metazoan cells. Most organisms form two microtubule arrays during mitosis, one oriented
toward the chromosomes to link each chromatid to its MTOC, and a second that assembles
between the two MTOCs to form a scaffold for MTOC separation. In G1 phase of the cell
cycle, prior to DNA and MTOC duplication, the primitive cell would have had a single MTOC,
with a single array of microtubules directed away from the nucleus that defined the polarity of
the cell. It is this cytoplasmic microtubule array that most likely provided the raw material for
evolution of the flagellar axoneme.42-44
6 Origins and Evolution of Eukaryotic Endomembranes and Cytoskeleton

Figure 3. Proposed steps in the transition from an early eukaryote, with a polarized morphology
based on asymmetric placement of a microtubule organizing center, but lacking flagella (left),
through an intermediate with a protoflagellum that supported gliding and limited bending (cen-
ter), to the last common eukaryotic ancestor, with a fully developed, motile 9+2 flagellum (right).

A polarized array of microtubules projecting from one side of the nucleus, as seen in most
cells today, need only become linked into a bundle to provide an organelle that could distort
the cell membrane and form a protoflagellum (Fig. 3). Microtubules radiating from the MTOC
that were not incorporated into this protoflagellar bundle would continue to provide a cytosk-
eleton for general cytoplasmic transport and organization of the endomembrane system, and
the interaction of motors such as kinesin and dynein with this microtubule cytoskeleton would
provide directed motility of associated vesicles. Such movement along the protoflagellar bundle
could direct exocytosis and endocytosis to a specific region of the cell membrane, creating a
new membrane domain. The similarity of IFT proteins to proteins involved in vesicle trafficking18
and the similarity of IFT kinesin and dynein to cytoplasmic versions of these motors, argues
that axonemes evolved from proteins that were already in use in microtubule-based vesicular
transport systems.
Early eukaryotes, lacking any other means of locomotion, were presumably benthic
amoeboid cells and could not yet swim. Microtubule-based motors moving along a parallel
bundle of microtubules in the protoflagellum would have provided at least two specific advan-
tages to these organisms. Simple coupling of retrograde movement to transmembrane proteins
would convert the protoflagellum to a feeding organelle, bringing particles that adhered to its
surface back toward the cell body by retrograde IFT for subsequent phagocytosis. Alternatively,
if substrate adhesion through IFT-associated protoflagellar transmembrane proteins was strong,
and cell body adhesion proportionately weak, retrograde IFT could support gliding motility.
Flagellar gliding as a means of locomotion is common in many pelagic, benthic and soil
flagellates today, and such surface motility has also been described for metazoan cilia, suggesting
that it was either an early adaptation of the IFT system, or one that has happened repeatedly
during subsequent evolution.44 As the coupling mechanisms between flagellar adhesion
molecules and gliding motors have not been widely studied, their evolutionary history remains
unknown.
Movement of secretory vesicles along a polarized microtubule bundle would also provide a
polarized distribution of cell surface molecules, including receptors. While the localization of
receptors to ciliary and flagellar surfaces has been documented in both protists and in meta-
zoan sensory cilia, including chemosensory cilia in C. elegans, kidney cilia in vertebrates, and
the highly modified cilia in sensory neurons of the vertebrate retina, evidence that specific
receptors have been localized to this membrane domain since before eukaryotic divergence is
only fragmentary (reviewed in ref. 44). Receptor localization does not require the complex
structure of a flagellum, but the ability to define a polarized cell surface domain for sensory
signaling may have been one of the driving forces in early flagellar evolution.
Flagella are first and foremost organelles that beat, and their complex structure would not
have evolved without strong selection for motility, as witnessed by the rapid loss of much of
The Evolution of Eukaryotic Cilia and Flagella as Motile and Sensory Organelles 7

this complexity in nonmotile sensory cilia, and the complete loss of flagella in nonmotile
organisms such as yeasts. However strong this advantage of a beating flagellum might have
been to early eukaryotes, intermediate stages must have existed that provided intermediate
levels of motion; a sudden jump from a benthic, amoeboid or gliding organism to one that can
swim by flagellar beating is not plausible. To understand the advantages of less vigorous
bending motility, one need only look at flagellar function among existing flagellates. Many
organisms use flagella to generate feeding currents that increase the frequency with which food
particles (bacteria, other eukaryotes, or detritus) can be ingested, while others use flagellar
movements to aid in trapping food particles.45,46 Feeding currents are common in organisms
such as choanoflagellates, which attach to the substrate with stalks and use flagellar beating to
create currents past the stationary cell, in mastigamoebae, which create currents while continuing
to move by amoeboid activity, and in many biflagellates such as bodonids, which create
currents with an anterior flagellum and glide on a posterior flagellum. Even a modest ability to
vibrate or wave a protoflagellum could have provided the initial selective advantage that drove
further development of single microtubules into doublet microtubules, and favored diversification
of dyneins that could take advantage of this new doublet microtubule track (Fig. 3).
Along with increased motility came the increased need to anchor the axoneme, which may
have driven both centriole/basal body evolution and the development of links between the
flagellar base and the cell membrane. These links would segregate the flagellar compartment
from the rest of the cytoplasm, requiring further refinements in the IFT sorting/trafficking
mechanism so that standard vesicle fusion occurred at the flagellar base, and IFT movement
transported both membrane and nonmembrane components to and from the flagellar
compartment. In addition, such links create a boundary between the flagellar membrane and
the rest of the plasma membrane, sequestering receptors and adhesion molecules into a unique
membrane domain.
The ability to bend does not require an axoneme with 9-fold symmetry, and many
axonemes have been discovered that depart from this pattern, yet most of these departures
appear to be more recent modifications of an ancestral 9+2 pattern. As argued in more detail
elsewhere, I propose that motility regulation by a central apparatus provided a strong selective
advantage to the organism in which it evolved, and that the most successful regulatory
mechanism was based on an apparatus built on a scaffold of two central microtubules, with
regulatory signals transmitted through radial spokes of a defined length.44,47 The geometry of
this regulatory mechanism presumably favors an outer cylinder of precisely nine doublet
microtubules, with the distance between doublets determined by the reach of dyneins that
must span each interdoublet gap. Whether central pair regulation was based on a fixed central
pair orientation, as found today in metazoans such as bivalves,48 sea urchins,49 and cteno-
phores50 and possibly in excavates such as euglenids,51 or a rotating central pair,52 as commonly
found in green algae such as Chlamydomonas53 and Micromonas,54 and chromalveolates such as
Paramecium55 and Synura,56 remains to be determined. Central pair rotation may allow regula-
tion of bends in different beat planes, and therefore be a more flexible regulatory system for
organisms whose survival is most highly dependent on rapid changes in flagellar beat param-
eters.47,53 The origin of radial spokes remains, at this time, one of the greater mysteries of
flagellar evolution, as related proteins have not been identified in other microtubule-associated
regulatory complexes.

Diversification of Flagellar Structure and Function

during Eukaryotic Radiation
Many changes to the basic (if complex) 9+2 flagellum that was present in the last common
eukaryotic ancestor are seen in some present day organisms, whereas others appear to have
retained the original model with few alterations. Changes include additions, such as
mastigonemes that project from the membrane surface and increase effective hydrodynamic
resistance, and accessory structures that increase axoneme stiffness (paraflagellar rods in
8 Origins and Evolution of Eukaryotic Endomembranes and Cytoskeleton

euglenoids, outer dense fibers or extra microtubules in spermatozoa; for a more extensive sur-
vey of structural changes in spermatozoa, see Baccetti, ref. 57). Simplifications or deletions of
structures no longer used by an organism or cell type include loss of the outer row dyneins, loss
of the central apparatus and radial spokes (motile 9+0), loss of the central apparatus, radial
spokes, and some doublet microtubules (6+0, 3+0). In the case of strictly nonmotile sensory
cilia, all components necessary for motility (central pair, spokes, dyneins) may be absent, leav-
ing only the membrane and the nine doublet microtubules to support IFT and receptor local-
ization. Motile flagella that lack the central pair and radial spokes appear to have a simplified
bending pattern that only accommodates helical bending waves, and are found on parasitic
flagellates that do not depend on flagellar motility for locomotion in a complex environment,58
on nodal cilia in early vertebrate embryos, where they generate a unidirectional fluid current
essential for establishing left-right asymmetry,59 and on certain vertebrate spermatozoa.60 Some
insect sperm that lack central pair microtubules have cylinders of 12 or 14 doublets, or spirals
of hundreds of doublets,57 suggesting further that the original standard of a cylinder of nine
doublet microtubules was selected to accommodate the radial spoke-central pair regulatory
complex, and that if the central pair is not needed, successful axonemes can evolve with alter-
native doublet patterns.
More difficult to catalog are modifications of existing parts to meet new demands, often
observable not at the structural level but as differences in average length, beat frequency, or
waveform, or as the ability to change beat frequency, beat direction, or waveform in response to
signaling cascades. These signaling pathways may in turn begin with stimulation of flagellar
surface receptors, or with cascades that are transmitted from elsewhere in the cell. The evolu-
tion of some of these changes, especially those affecting signaling pathways, will be difficult to
trace until many more genomes have been sequenced, and proteomic analysis confirms the
location and function of putative flagellar gene products. Some changes probably occurred
only once, and surveying the distribution of organisms that retain such features may clarify
phylogenetic relationships. Other changes have occurred independently more than once and
can be considered convergent evolution. For example, the ability to generate ATP within the
flagellar compartment has evolved in several independent ways. Glycolysis is an important
source of energy for sperm motility in mammals61 where fermentable sugars come directly
from seminal fluid,61,62 but has not been reported in vertebrate cilia and flagella other than
sperm tails and some types of nonmotile cilia (e.g., the outer segments of mammalian photore-
ceptor cells63). Mammalian sperm-specific isoforms of glycolytic enzymes such as glyceralde-
hyde 3-phosphate dehydrogenase64 and enolase65 have likely evolved recently, as the glycolytic
enzymes identified in Chlamydomonas flagella66 are not closely related and appear to have been
targeted for flagellar use specifically in algae. Completely different methods of flagellar ATP
generation occur in other organisms, including phosphocreatine/creatine phosphokinase shuttles
in sea urchin67 and mammalian68 spermatozoa, and in chicken photoreceptor outer segments,69
and a phosphoarginine/arginine phosphokinase shuttle in Paramecium cilia.70 Overall, the ba-
sic motile machinery of 9+2 organelles appears to be ancient and highly conserved, whereas the
signaling cascades that regulate motility and accessory structures that modify its output have
changed to suit specific organismal needs.

Summary
Typical 9+2 flagella likely evolved from bundling and extension of cytoplasmic microtubules
that assembled on a microtubule organizing center and that generated a polarized cellular
morphology. Close apposition of the plasma membrane created a separate membrane domain
that could be used to localize receptors for sensory signal transduction, and required
simultaneous evolution of intraflagellar transport to maintain this polarized structure.
Membrane-associated IFT-based movement provided a mechanism for gliding motility, and
addition of axonemal dynein motors allowed this extension to bend and generate currents past
the cell. Formation of doublet microtubules allowed elaboration of dyneins to improve
The Evolution of Eukaryotic Cilia and Flagella as Motile and Sensory Organelles 9

motility, and the addition of the radial spoke-central pair regulatory system provided responsive
dynein control. The strong selective advantage of a motile 9+2 flagellum may have resulted in
rapid diversification of the last common eukaryotic ancestor into all existing branches of
eukaryotic organisms.

Acknowledgements
Robert Bloodgood contributed through thoughtful discussions on flagellar gliding, David
Asai shared recent results on dynein evolution, and Gaspar Jekely provided insight into the
evolutionary connections between IFT and vesicle trafficking.

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Correction:

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