The Serial Endosymbiosis Theory of Eukaryotic Evolution

by
Jeremy Mohn

(c) 1998

The transition between eukaryotes, cells with nuclei, and prokaryotes, cells which lack
nuclei, is considered by many biologists to be the most profound change in evolutionary
history. In an attempt to describe the way in which this gap was bridged, scientists have
proposed the serial endosymbiosis theory (SET). The term "endosymbiosis" specifies the
relationship between organisms which live one within another (symbiont within host) in a
mutually beneficial relationship. The SET states that the evolution of eukaryotes from
prokaryotes involved the symbiotic union of several previously independent ancestors.
According to the theory, these ancestors included a host cell, an ancestor of mitochondria, an
ancestor of chloroplasts, and, more controversially, a prokaryote that brought with it the
structures that today provide cellular motion.

The idea that the eukaryotic cell is actually a colony of microbes was first suggested in the
1920s by American biologist Ivan Wallin (Fausto-Sterling 1993). The originator of the
modern version of the SET is biologist Lynn Margulis. In 1981, Margulis published the first
edition of her book entitled Symbiosis in Cell Evolution in which she proposed that
eukaryotic cells originated as communities of interacting entities that joined together in a
specific order. With time, the members of this union became the organelles of a single host
(Margulis 1993). The organelle progenitors could have gained entry into a host cell as
undigested prey or as an internal parasite after which the combination became mutually
beneficial to both organisms. As the organisms became more interdependent, an obligatory
symbiosis evolved.

The SET postulates that the ancestors of mitochondria were free-living bacteria, similar to
today’s Daptobacter and Bdellovibrio, that developed the ability to efficiently respire oxygen
(Margulis and Sagan 1987). The ancestors of chloroplasts, today’s cyanobacteria, were
originally independent photosynthetic organisms. In addition, the whiplike cilia that are
common in eukaryotes but are not found in prokaryotes are thought to have derived from still
another group of free-living bacteria, the modern spirochetes. According to the SET, the
original prokaryotic host cell was an archaebacterium, similar to today’s Thermoplasma, that
could withstand high temperatures and acidic conditions (Margulis and Sagan 1987). This
host cell was neither photosynthetic nor capable of effectively using oxygen.

Throughout her writings, Margulis contends that symbiosis is a major driving force behind
evolution. In her opinion, cooperation, interaction, and mutual dependence among life forms
allowed for life’s eventual global dominance. As a result, Darwin’s notion of evolution as the
"survival of the fittest," a continual competition among individuals and species, is
incomplete. According to Margulis and Sagan (1986), "Life did not take over the globe by
combat, but by networking." Rather than focus solely on the elimination of competitors,
Margulis’ view of evolution downplays competition itself on the basis of symbiotic
relationships.

One early and important discovery in support of the SET occurred in the laboratory of
Kwang W. Jeon, a biologist at the University of Tennessee. Jeon witnessed the establishment
of an amoeba-bacteria symbiosis in which new bacterial symbionts became integrated in the
host amoeba (Jeon 1991). In 1966, when the bacteria first infected the amoebas, they were
lethal to their hosts. However, as time progressed, some of the infected amoebas survived and
became dependent on their newly acquired endosymbionts within a few years. Jeon was able
to prove this dependency by performing nuclei transplants between infected amoebas and
amoebas lacking the bacteria. If left alone, the hybrid amoebas died in a matter of days. Yet if
he reinfected these hybrids with the once-lethal bacteria, the amoebas recovered and once
again began to grow (Margulis and Sagan 1987). This discovery served to demonstrate that
endosymbiosis could provide a major mechanism for cellular evolution and explain the
introduction of new species (Jeon 1991).

Although some of Margulis’ ideas remain controversial, there is mounting evidence in

support of the SET (Fausto-Sterling 1993). The bulk of this evidence serves to defend the
notion of an endosymbiotic origin of mitochondria and chloroplasts. The recognition that new
mitochondria and chloroplasts can arise only from preexisting mitochondria and chloroplasts
was one of the first clues. Scientists found that mitochondria and chloroplasts cannot be
formed in a cell that lacks them because nuclear genes only code for some of the proteins of
which they are made. Also, both mitochondria and chloroplasts have their own sets of genes
that are more similar to those of prokaryotes than those of eukaryotes. They both contain a
circular molecule of DNA, just like that found in prokaryotes. Finally, both mitochondria and
chloroplasts have their own protein-synthesizing machinery. Their ribosomal structures and
their ribosomal RNA (rRNA) more closely resemble those of prokaryotes. These three lines
of evidence have been cited to firmly establish the theory of the origin of mitochondria and
chloroplasts through the process of endosymbiosis.

The least accepted and most questionable aspect of the SET is the hypothesis that
eukaryotic undulipodia originated from spirochete bacteria (Margulis 1993). The term
"undulipodia" is used to describe the eukaryotic motility organelles, flagella and cilia.
Undulipodia are composed of microtubules in a specific configuration. Microtubules are
comprised of several closely related proteins called tubulins. These structures are far larger
and more complex than bacterial flagella, which are made of flagellin proteins. The SET
postulates that undulipodia may be derived from bacteria through motility symbioses
(Bermudes, Margulis, and Tzertzinis 1987). This idea is referred to as the exogenous
hypothesis. The details of the argument are complex, but the supporters of the SET point to
several lines of circumstantial evidence. Their argument emphasizes the biology of the
organelles themselves, their distribution, and the occurrence of related and analogous
structures. Opponents of this view, supporters of the endogenous hypothesis, suggest that
undulipodia originated internally as an extension of the microtubules utilized in mitosis. This
hypothesis is also referred to as direct filiation, which is the nonsymbiotic view of evolution
that emphasizes the role of various kinds of mutations in the evolutionary separation of
eukaryotic cells from prokaryotic cells.

The main controversy between the endogenous and exogenous hypotheses for the origin of
undulipodia rests upon a question of chronology. Proponents of the endogenous hypothesis
claim that microtubules preceded the origin of undulipodia, which eventually arose
endogenously. In contrast, the exogenous hypothesis states that motility symbioses gave rise
to cells with undulipodia, and this acquisition subsequently led to the internal structures
involved in mitosis (Bermudes, Margulis, and Tzertzinis1987). Although the symbiotic origin
of undulipodia is gaining support, the controversy is yet to be solved. According to Bermudes
and Margulis (1985), there is insufficient evidence to prove either direct filiation or the
symbiotic hypothesis for the origin of undulipodia.

An important distinctive element of the SET is the overall chronology of symbiotic

acquisitions in the origin of the eukaryotic cell. In order to fully understand the theory’s
implications for the classification of all life forms, a brief summary of the current
interpretation of endosymbiotic events is necessary. According to the theory, eukaryotes
evolved when archaeal and eubacterial cells merged in anaerobic symbiosis. The archaeal cell
provided the cytoplasm while the eubacterial cell (a spirochete) allowed for mobility and,
eventually, mitosis. Some of these anaerobic cells then incorporated oxygen-respiring
eubacteria (similar to Daptobacter or Bdellovibrio) to become mitochondria-containing
aerobes from which most protoctists, animals, and fungi evolved. Finally, some of these
aerobes went on to incorporate photosynthesizing cyanobacteria to become chloroplast-
containing algae and plants. The divisions or domains implied by this description (Archaea,
(true)Bacteria, and Eukarya) are consistent with the widely acknowledged classification
system described by Olsen, Woese, and Overbeek (1994).

Although the nucleus is the defining characteristic of the eukaryotic cell, the origin of this
organelle and its relation to symbiosis is uncertain. Margulis tends to favor a process
involving the combination of direct filiation and symbiosis as the source of the nucleated cell.
She believes that some prokaryotic cells evolved primitive nuclei through direct filiation but
remained prokaryotic. Others evolved these same structures but also acquired other symbiotic
genes and consequently became eukaryotes (Margulis 1993). Overall, the traditional view of
the origin of the nucleus states that the nuclear genome originated through direct evolution
from an archaebacterial ancestor.

A 1996 paper by Golding and Gupta disputes the traditional view of the origin of the
nucleus and suggests an alternative called the chimeric model. The term "chimeric" refers to
an organism containing tissues from at least two genetically distinct parents. The chimeric
model proposes that the first eukaryotic cell arose as the result of a unusual fusion event
between a Gram-negative eubacterium (host) without a cell wall and an archaebacterium
(symbiont) in which both parents made major contributions to the cell’s nuclear genome. The
nucleus appeared as the result of the folding in of the host’s membrane around the engulfed
cell. Such fusion events are generally rejected by supporters of the SET because of the
inability of present-day bacteria to envelope prey.
The chimeric model is based on genetic and biochemical evidence. One piece of genetic
evidence that supports the model is the fact that prokaryotic cells are homogenomic (having
genetic material from one parent only), whereas eukaryotic cells are heterogenomic (having
genetic material from more than one parent). Biochemical evidence in support of the chimeric
model involved the phylogenetic, or evolutionary, analysis of sequence data from proteins.
This analysis demonstrated a close relationship between Gram-negative bacteria and
eukaryotes on one hand and Gram-positive bacteria and archaebacteria on the other (Golding
and Gupta 1996). Even more protein sequence data suggested a relationship between
eukaryotes and archaebacteria. These data imply that a symbiotic relationship between Gram-
negative bacteria and archaebacteria as the progenitors of the eukaryotic cell is feasible.
Overall, the sequence data support the chimeric model.

Recent research by Martin and Müller (1998) into the origin of the mitochondrion has led to
a new theory of endosymbiosis called the "hydrogen hypothesis." In the current picture of the
origin of the eukaryotic cell, the mitochondrion was a "lucky accident" (Vogel 1998). The
ancestral host cell simply engulfed the mitochondrion ancestor, did not fully ingest it, and an
even more successful cell resulted. According to the hydrogen hypothesis, however, the first
eukaryotic cell did not form simply by accident. Instead, it was the result of a purposeful
union between an archaebacterial host cell, a methanogen that consumed hydrogen and
carbon dioxide to produce methane, and a future mitochondrion symbiont that made
hydrogen and carbon dioxide as waste products of anaerobic metabolism. Thus, although the
symbiont was probably capable of aerobic respiration, the symbiosis began as a result of the
products of anaerobic metabolism. The host’s dependence upon hydrogen produced by the
symbiont is identified as the selective principle that consolidated the common ancestor of
eukaryotic cells (Martin and Müller 1998).

The hydrogen hypothesis has some important implications that contradict the current view
of the relationship between eukaryotes and archaebacteria. In the current view, the eukaryotes
branched off from the archaebacteria long before the archaebacteria had divided into their
present-day groups. The hydrogen hypothesis implies that the first eukaryotes appeared much
later in the evolutionary picture, meaning they are more closely tied to the archaebacteria. In
order for the hydrogen hypothesis to be confirmed, an analysis of the complete sequences of
eukaryotic and archaebacterial genomes must be completed (Vogel 1998).

Another recent explanation of the origin of eukaryotes called the "syntrophic hypothesis"
was presented by López-García and Moreira (1998). Though they were independently
proposed, the syntrophic hypothesis is complementary in several aspects to the hydrogen
hypothesis. Both hypotheses agree in the suggestion of an anaerobic metabolism for the
origin of mitochondrial symbiosis. They are also strikingly similar in some metabolic details
of the symbiosis and archaeal molecular features (López-Garcia and Moreira 1998). The
major difference between the two hypotheses is in the nature of the original bacterial
partnership. As previously stated, in the hydrogen hypothesis, the original symbiosis is
thought to have taken place between a methanogenic archaebacterium and a eubacterial
ancestor to the mitochondrion. In the syntrophic hypothesis, the original symbiosis is
conceived to have taken place between a methanogenic archaebacterium and an ancestral
sulfate-respiring delta-proteobacterium. The former provided the central genetic material and
nucleic acid metabolism while the latter provided most metabolic characteristics (López-
Garcia and Moreira 1998). Mitochondria are thought to have derived from a later,
independent symbiotic event. As with the hydrogen hypothesis, further genetic sequencing
analyses are necessary in order for the claims of the syntrophic hypothesis to be upheld.

It has been nearly thirty years since Lynn Margulis first published a book on the origin of
eukaryotic cells. Since that time, biology has undergone extraordinary changes. The most
noticeable change is the extensive accumulation of sequence data for both nucleic acids and
proteins. The collection of new data will undoubtedly lead to continuous revision of the serial
endosymbiosis theory of the origin of the eukaryotic cell. Despite the uncertain future, the
crucial foundation has been laid. Symbiosis is now accepted by the scientific community as
an important factor in generating evolutionary change. The next steps include the
development of more elaborate methods to interpret genetic and molecular sequence data and
the undertaking of a fresh look at the fossil record. These tactics might reveal significant
information concerning one of the most challenging and fascinating problems in evolutionary
biology, the origin of the eukaryotes.

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