Arch Microbiol (1996) 166 : 283–292
MINI-REVIEW
Arthur L. Kruckeberg
The hexose transporter family of Saccharomyces cerevisiae
Received: 24 June 1996 / Accepted: 29 July 1996
Abstract Saccharomyces cerevisiae accomplishes high
rates of hexose transport. The kinetics of hexose transport
are complex. The capacity and kinetic complexity of hex-
ose transport in yeast are reflected in the large number of
sugar transporter genes in the genome. Twenty hexose
transporter genes exist in S. cerevisiae. Some of these
have been found by genetic means; many have been dis-
covered by the comprehensive sequencing of the yeast
genome. This review codifies the nomenclature of the
hexose transporter genes and describes the sequence ho-
mology and structural similarity of the proteins they en-
code. Information about the expression and function of
the transporters is presented. Access to the sequences of
the genes and proteins at three sequence databases is pro-
vided via the World Wide Web.
Key words Hexose transport · Glucose transport ·
Galactose transport · Genome sequence analysis ·
Telomere
The obligate first step of sugar metabolism is sugar trans-
port. In the yeast Saccharomyces cerevisiae, hexoses are
delivered to cellular metabolism by facilitated diffusion
across the plasma membrane. The rate of transport by
cells that are fermenting rapidly can exceed 107 glucose
molecules per cell per second. Transport has been inferred
to be the rate-limiting step of glycolysis (Becker and Betz
Dedicated in memory of Professor Michael Ciriacy
Supplementary material Further details (Tables W1–W3 and
Figs. W1–W4) have been deposited in electronic form and can be
obtained from http://science.springer.de/archmicro/am.htm
A. L. Kruckeberg
Department of Biological Chemistry, UCLA School of Medicine,
CHS 33–257, 10833 Le Conte Avenue, Los Angeles,
CA 90095–1737, USA
Tel. +1-310-825-8363; Fax +1-310-206-5272
e-mail: kruckeba@biovx1.biology.ucla.edu
© Springer-Verlag 1996
1972; Gancedo and Serrano 1989); in the description of
metabolic control analysis (Fell 1992), transport has a
high degree of control on glycolytic flux (Cortassa and
Aon 1993; Oehlen et al. 1994).
The kinetic characteristics of hexose transport have been
characterized by many investigators. However, the mole-
cular basis of hexose transport is only beginning to be un-
derstood. This is due in part to the intractibility of the hy-
drophobic transport proteins to biochemical analysis, and in
part to the genetic complexity of the hexose transport sys-
tem in yeast. Now, with the complete sequence of the S.
cerevisiae genome available for study, all of the candidate
genes for the hexose transporters have been identified. The
number of such candidates is surprisingly large. The roles
of the plethora of sugar transporters is a fundamentally im-
portant question in yeast biology. Do they differ in their ki-
netic or functional characteristics? Are they differentially
regulated in time or space? Or are they redundant, and is their
number a consequence of genomic instability? The purpose
of this review is to enumerate the hexose transporter gene
family in yeast and to comment on some of the distinctive
features of those genes and the proteins they encode.
This review is intended to be topical rather than com-
prehensive. Readers are referred to recent reviews on
sugar transport in yeast (Bisson et al. 1993; Ciriacy and
Reifenberger 1996; Lagunas 1993; Weusthuis et al. 1994)
for more detailed discussions. Preliminary reports on the
complexity of the HXT family of sugar transporter genes
have been published by André (1995) and by Goffeau and
co-workers (Nelissen et al. 1995). The tables and figures
presented in this text are supplemented by tables and fig-
ures that are available via World Wide Web (http://sci-
ence.springer.de). Links from this server will lead the in-
terested reader to the sequences of the genes and proteins
at the Saccharomyces Genome Database, the Martinsried
Institute of Protein Science, and the Yeast Protein Data-
base. References to figure and table numbers of the sup-
plementary materials available via World Wide Web are
indicated with a “W.”
284
Background
The natural habitat of S. cerevisiae is fruit juices. This en-
vironment is rich in solutes, particularly sugars. As a con-
sequence of growth and fermentation of these sugars,
yeast brings about dramatic changes in its physicochemi-
cal environment, and to sustain its growth it must adapt to
these changes. The sugar concentration may decline from
1 M to 10–5 M during a fermentation, and the overall com-
position of the medium will be altered by cellular metab-
olism. The sugar transport activity of the cell and the pro-
teins that mediate sugar transport must be responsive to
these changing conditions.
S. cerevisiae transports hexoses via facilitated diffu-
sion. Transport occurs with low affinity (Km ≈ 20 mM for
glucose) in cells grown in glucose-rich media. An addi-
tional high affinity component (Km ≈ 1 mM for glucose) is
expressed in cells grown in media with limiting hexose
concentrations (see Coons et al. 1995; Gamo et al. 1995
and references therein). In both batch fermentations and
chemostat cultures, the kinetics of glucose transport are
modulated: high-affinity transport is expressed as a greater
component of the total uptake capacity at later stages of
batch fermentations or at low dilution rates, i.e., in condi-
tions in which the glucose concentration is limiting
(Postma et al. 1989; Walsh et al. 1994). In other words,
yeast adapts to the concentration of hexoses in its envi-
ronment by expressing a transport system that has a sub-
strate affinity appropriate to those conditions.
The previous paragraph is a rather cursory discussion
of the large body of work from many laboratories on
the kinetics of hexose transport in yeast. Moreover, it
skirts the controversies that exist in the field. However,
since the powers of molecular biology can finally be
brought fully to bear on sugar transport in yeast, it can be
anticipated that these controversies will be resolved and
that the kinetics of transport can be explained mechanisti-
cally.
Hexose transporter genes have been identified by three
means in S. cerevisiae. Some of the genes have been iso-
lated in genetic studies. The first gene isolated was SNF3.
Mutants were isolated that were defective in growth on
sucrose and raffinose (Neigeborn and Carlson 1984).
Most of these were defective in derepression of SUC2
(encoding extracellular invertase) when grown on sucrose
or raffinose. However, the snf3 mutants were shown to be
impaired in uptake of the low levels of hexoses produced
by hydrolysis of these sugars and in high affinity glucose
transport activity (Bisson et al. 1987). The cloned SNF3
gene was found to encode a hydrophobic protein with ho-
mology to sugar transporters from mammals and bacteria
(Celenza et al. 1988). Soon afterwards, the galactose
(GAL2; Nehlin et al. 1989; Szkutnicka et al. 1989) and
maltose (MAL61; Cheng and Michels 1990; Yao et al.
1989) transporter genes were shown to encode proteins
homologous to Snf3. Subsequently, genes that suppress
the growth defect of a snf3 null mutation when supplied in
multi-copy were shown to encode Snf3 homologs as well
(Kruckeberg and Bisson 1990; Lewis and Bisson 1991;
Theodoris et al. 1994). These genes, HXT1, HXT2, and
HXT4, were named HXT for hexose transport. They have
also been recovered as multicopy suppressors of a domi-
nant mutation in the HTR1 gene; HTR1 is involved in hex-
ose transport regulation (Özcan et al. 1993).
Hexose transporters have been identified in other ge-
netic screens as well. For example, mutations in HXT1,
HXT3 and HXT4 (Ko et al. 1993) as well as various other
solute transporter genes (R. Gaber, personal communica-
tion) are able to suppress mutations in the potassium trans-
porter genes, presumably because the mutant solute trans-
porters have lost the ability to exclude cations from their
transport duct. The HXT4 (LGT1; Prior et al. 1993; Gen-
Bank X67321) and HXT11 (LGT3; GenBank X82621)
genes were cloned by their ability to complement a muta-
tion in the RAG1 low-affinity glucose transporter gene of
Kluyveromyces lactis.
HXT1 was recovered as one of a set of cDNAs repre-
senting genes that are differentially expressed in hyperos-
motic media; cDNAs encoding glucokinase and glycerol-
3-phosphate dehydrogenase were also found in this set.
This suggests that HXT1 expression is elevated in hyper-
osmotic conditions to provide the substrate for synthesis
of the osmoprotectant glycerol (Hirayama et al. 1995).
The HXT13 promoter was found in a collection of pro-
moters that are regulated by the Hap2 transcriptional reg-
ulator and are repressed by glucose (B. Daignan-Fornier,
personal communication); this suggests that the HXT13
gene is expressed in glucose-limited conditions coordi-
nately with respiratory functions.
The remaining HXT genes have been discovered mostly
by direct means. HXT5, HXT6 and HXT7 were isolated by
DNA hybridization screening with the HXT3 gene (Ciri-
acy and Reifenberger 1996; Reifenberger et al. 1995). The
remainder have been discovered in the course of the sys-
tematic sequencing of the yeast genome.
The hexose transporter gene family: sequence similarity
The hexose transporter family in yeast, as defined here,
includes 20 genes. These genes are listed in Table 1, and
links to their DNA and protein sequences in the databases
are provided in Table W1. The family has been defined on
the basis of sequence similarity among the proteins they
encode (Fig. 1) and on the basis of related function. The
HXT genes have been named according to a convention
agreed upon by researchers in the field of hexose trans-
porter molecular genetics and accepted by the Saccha-
romyces Genome Database (L. Bisson, R. Gaber, M. Ciri-
acy, M. Cherry, personal communications). The SNF3 and
RGT2 genes delimit this family; they have significant se-
quence similarity with HXT1 – HXT17 and GAL2, and they
have hexose transport phenotypes. GAL2 and some of the
HXT genes are known to have hexose transport pheno-
types as well; most of the others have not been investi-
gated. As Saier has pointed out, sequence similarity corre-
lates well with substrate specificity among transport pro-

Fig. 1 Dendrogram of sequence similarity among yeast hexose
transport proteins. The dendrogram shows the clustering relation-
ships of the sequences based on their similarity; the length of the
horizontal branches is inversely proportional to the sequence simi-
larity of the sequences at each branch tip. The sequences were
clustered by the GCG PILEUP program, and the dendrogram was
plotted by the GCG FIGURE program (Genetics Computer Group
1994). The chromosomal location of the gene encoding each trans-
porter is shown next to the protein name
teins (Saier 1994). Therefore, it is reasonable to consider
all of these genes as a family.
The proteins range from 99.7% identical (Hxt6 and
Hxt7 differ by only a single, conserved amino acid) to
25% identical (Snf3 with Hxt16 or Hxt17) (Table W2).
The similarities range from 46.9% (Rgt2 and Hxt5) up to
99.7% (Hxt6 and Hxt7). The proteins are similar in size as
well, ranging in length from 540 to 592 residues; two im-
285
portant exceptions to this are the Rgt2 and Snf3 proteins,
which are 200 and 300 residues longer than the others, re-
spectively.
The hexose transporters comprise a subfamily: a num-
ber of other genes have been identified in yeast that encode
homologs of these proteins. However, these homologs
mediate transport of other solutes (myo-inositol, phosphate,
disaccharides, or unknown solutes) and have considerably
lower sequence similarity to the Hxt proteins than the sub-
family members have to one another (Bisson et al. 1993).
The subfamily is also homologous to a much larger fam-
ily of sugar transporters identified in plants, animals and
bacteria, and to transporters of other solutes in these king-
doms and in fungi as well (Griffith et al. 1992). The se-
quence relationships among representative proteins of the
sugar transporter superfamily is diagrammed in Fig. W1.
For the sake of comparison, the similarity of members
of the yeast hexose transporter family to other yeast trans-
porters is: the Itr1 inositol transporter, 38–46% similarity;
the Mal61 maltose transporter, 36–43% similarity; and the
Pho84 phosphate transporter, 35–42% similarity. The sim-
ilarity of the hexose transporter family members to other
hexose transporters is: the human GLUT1 glucose trans-
porter, 42–48% similarity; the Arabidopsis thaliana Stp1
glucose-H+ cotransporter, 43–49% similarity; the galP
galactose transporter of E. coli, 46–53% similarity. The
Hxt transporters have 45–51% similarity to the qa-Y
quinate transporter of Aspergillus nidulans.
Despite the low absolute values of sequence similarity
between some of these transporters, they are definitely ho-
mologous (i.e., the genes are descended from a common
ancestral gene) on the grounds that they share highly con-
served sequence motifs. They are also homologous on the
basis of the transitivity test (Pearson 1996): for example,
Mal61 and Pho84 are only 12% identical, which is not
statistically significant by the shuffle test (Saier 1994), but
each is 19% identical with Hxt13, and this similarity is sig-
nificant (data not shown; Bisson et al. 1993). Since Pho84
and Mal61 are each homologous with Hxt13, then they must
be homologous with each other. The homology of the
yeast hexose transporters with the other transporters de-
scribed above has been shown previously for the gene se-
quences available at that time (Bisson et al. 1993). Due to
the homology among all of the yeast hexose transporters,
the homology of the recently identified Hxts to the sugar
transporter superfamily is also inferred by transitivity.
The sequences of the 20 proteins in the hexose trans-
porter family have been aligned; a plot of the degree of
similarity is shown in Fig. 2, and the multiple sequence
alignment is presented in Fig. W2. The similarity plot re-
veals that the proteins are conserved throughout most of
their length. The unusual size of the Rgt2 and Snf3 pro-
teins is due to unique sequences at their carboxyl termini
(Fig. W2). These carboxy-terminal sequences have a highly
conserved core sequence that is present once in Rgt2 and
is duplicated in tandem in Snf3. The core sequence is not
encoded elsewhere in the yeast genome.
A number of regions are particularly highly conserved
among the 20 hexose transporters. Two sugar transporter
286
Fig. 2 The average similarity at each position of the multiple se-
quence alignment of the yeast hexose transporters was calculated
with a comparison window of 12, using the GCG PLOTSIMI-
LARITY program. The average similarity for the whole alignment
is shown as a dotted line. The predicted positions of the trans-
membrane domains are shown as black rectangles above the plot;
these were estimated using the TopPred II (Claros and von Heijne
1994) and TMAP (Persson and Argos 1994) topology prediction
programs
signature sequences are presented in the PROSITE dictio-
nary of protein motifs (Bairoch 1993); the yeast hexose
transporter sequences have both of them. The consensus
sequences of the motifs in these proteins are:
i
– vl s migration is not affected by treating cells with the glyco-
v sGlGvGgia––
av– s Pmli a– EvapkhlR
(starting at position 223 of the consensus sequence, Fig.
W3; lowercase indicates that the residues are not abso-
lutely conserved at this position, and x z– means that only
residues x and z are present at this position).
v
––d– rfG ––––
r r c LlwG
ie h kv
(starting at position 442).
Another highly conserved motif is
l ryy
– PESP ––––
v qf l
(starting at position 307).
These highly conserved motifs occur in all members of
the sugar transporter family from bacteria to fungi, plants,
and animals.
Fifty-five residues are absolutely conserved in the 640
residue region of the multiple sequence alignment preced-
ing the Snf3 and Rgt2 carboxy-terminal region. By far the
most highly conserved residue is glycine (18 occur-
rences), followed by proline and glutamate (5 each) and
phenylalanine (4).
Asparagine residues may be glycosylated if they occur
in the sequence NX — S and are exposed to the lumen of the
T (Fig. 3). The region of this domain in these proteins con-
endoplasmic reticulum. A potential asparaginyl glycosy-
lation site occurs in a conserved location at position 128
of the consensus sequence (Fig. W3; e.g., residue 82 of
Hxt2, Fig. W2). It has the sequence NQT in Hxt2, Hxt6,
Hxt7 and Hxt8, and NLS in Hxt9, Hxt11 and Hxt12. Hxt8
has a second site (NYS) nine residues towards the car-
boxyl terminus from the first, and Hxt5 has an NGT site
three residues further away. Topological models of these
proteins suggest that this region would be exposed to the
ER lumen en route to the plasma membrane (see below).
The mammalian GLUT1 glucose transporter is glycosy-
lated at an NQT sequence that occurs in a topologically
identical location in the protein (Mueckler et al. 1985). It
is not known whether any of the yeast hexose transporters
are glycosylated. Hxt2 migrates as a diffuse band in SDS-
PAGE gels, which is indicative of glycosylation, but its
sylation inhibitor tunicamycin during expression of Hxt2
(Wendell and Bisson 1993).
Secondary structure
The secondary structure of the hexose transport proteins is
highly conserved. Figure 3 shows the Chou-Fasman hy-
drophobicity profiles of two representative members of
the family, Hxt5 and Hxt14. The hydrophobicity profiles
of the other 18 members are presented in Fig. W4. The
proteins have a number of highly hydrophobic regions, in-
terspersed with regions of low hydrophobicity or net hy-
drophilicity. The amino and carboxyl termini are hydro-
philic. When the topologies of the proteins are predicted
by the TopPred II topology prediction program (Claros
and von Heijne 1994), 11 or 12 discrete hydrophobic do-
mains are demarcated that are potential transmembrane α-
helices (Table W3). Two pairs of putative transmembrane
domains (1 and 2, and 6 and 7) are separated by particu-
larly long hydrophilic regions. The amino and carboxyl
termini are predicted to be on the cytosolic side of the
membrane.
When sequences with 12 predicted transmembrane do-
mains are compared with sequences that are predicted to
have 11 domains, the 7th domain is missing in the latter
tains a few more hydrophilic residues and does not reach
the threshold hydrophobicity required to specify it as a

Fig. 3A, B Hydrophobicity
plots of two yeast hexose
transport proteins. A Hxt5 is
predicted to have 11 trans-
membrane domains. B Hxt14
is predicted to have 12 trans-
membrane domains. The plots
were constructed from the hy-
drophobicities determined at
each residue by the method of
Kyte and Doolittle (1982) us-
ing GCG PEPPLOT with an
averaging window of 21
residues. Hydrophobic regions
are numbered
transmembrane domain. It has been pointed out that seg-
ments of proteins can span the lipid bilayer without being
highly hydrophobic if they are largely in contact with
other more hydrophobic membrane-spanning segments
(Lodish 1988). From this point of view it is likely that all
of the yeast hexose transport proteins have 12 domains
that traverse the membrane. This secondary structure has
also been predicted for all of the homologs of these pro-
teins. The structural model for the mammalian hexose
transporter proteins has been described elsewhere (e.g.,
Baldwin and Henderson 1989; Gould and Holman 1993).
The secondary structure of only one of these, the human
erythrocyte glucose transporter GLUT1, has been studied
in detail. Biochemical and immunochemical analyses of
GLUT1 have shown that the amino and carboxyl termini
are directed towards the cytoplasm and are in agreement
with the prediction of 12 transmembrane domains. More
recently, the topology of GLUT1 was characterized by in-
sertion via mutagenesis of a glycosylation site into each
hydrophilic region of the protein. When the mutant proteins
were expressed in Xenopus oocytes, those with glycosyla-
tion sites inserted into exofacial loops (as predicted by the
model) were in fact glycosylated, while those with glyco-
sylation sites that were predicted to be oriented towards
the cytoplasm were not glycosylated (Hresko et al. 1994).
The consensus positions of the 12 putative transmem-
brane domains in the multiple sequence alignment of the
yeast hexose transporters have been estimated using both
the TopPred II (Claros and von Heijne 1994) and TMAP
(Persson and Argos 1994) topology prediction programs,
and are shown above the similarity plot in Fig. 2. The re-
gions of highest sequence conservation among the Hxt
proteins are found in or adjacent to the transmembrane
domains, especially domains 1, 2, 4, 6, 7, 11, and 12.
287
Most of the gaps that have been introduced to align the se-
quences occur outside of the membrane-spanning regions.
This relationship between sequence conservation and sec-
ondary structure is reasonable since the transmembrane
domains are expected to have conserved structural require-
ments. Secondary structure prediction (Mueckler et al. 1985)
and biochemical and mutational analyses (e.g., Gould and
Holman 1993; Mueckler et al. 1994) have implicated do-
mains 3, 5, 7, 8, and 11 as being involved with substrate
binding or forming the hydrophilic duct through which
sugars are translocated across the membrane. This is sup-
ported by the analysis of chimeras between Gal2 and Hxt2;
the analysis localizes the substrate recognition region to
the last three transmembrane domains of Gal2 and a por-
tion of the carboxy-terminal tail (Nishizawa et al. 1995).
The arrangement of the 12 transmembrane domains of
the hexose transport proteins in a 3-dimensional hexose-
transporting structure is a fundamental issue in under-
standing the function of the transporters. A tertiary struc-
ture has not yet been determined for any member of the
sugar transporter superfamily, though a molecular model
has been developed for GLUT1 (Gould and Holman 1993).
Genomic distribution and evolution
From Table 1 and Fig. 1 it can be seen that the 20 mem-
bers of the hexose transporter gene family are located on
10 of the 16 chromosomes of yeast. Some clues on the
evolution of the family can be discerned by examining the
distribution of the genes in the genome. HXT2 and HXT10
are closely related in sequence, and both of these genes
are centromere-linked. Eight of the HXT genes are in or
adjacent to the subtelomeric regions of their chromosomes.
288
Table 1 The members of the Saccharomyces cerevisiae hexose transporter gene family. L left arm, R right arm, C centromere proximal,
ST in or adjacent to the subtelomeric region
Genea Number of codons Codon Adaptation Indexb Chromosome Open reading frame
HXT1 570 0.408 VIII-R YHR094c
HXT2 541 0.359 XIII-R C YMR011w
HXT3 567 0.494 IV-R YDR345c
HXT4 576c 0.551 VIII-R YHR092c
HXT5 592 0.206 VIII-R YHR096c
HXT6 570 0.519 IV-R YDR343c
HXT7 570 0.519 IV-R YDR342c
HXT8 569 0.202 X-L ST YJL214w
HXT9 569 0.167 X-L ST YJL219w
HXT10 546 0.164 VI-L C YFL011w
HXT11 567 0.166 XV-L ST YOL156w
HXT12 567d 0.173 IX-L ST YIL170w + YIL171w
HXT13 564 0.193 V-L ST YEL069c
HXT14 540e 0.107 XIV-L YNL318c
HXT15 567 0.162 IV-L ST YDL245c
HXT16 567 0.162 X-R ST YJR158w
HXT17 564 0.201 XIV-R ST YNR072w
GAL2 574 0.245 XII-R YLR081w
SNF3 884 0.130 IV-L YDL194w
RGT2 763 0.119 IV-L YDL138w
a The DNA sequences and locations were obtained from the Yeast d ORFs YIL170w and YIL171w have been combined by deletion
Genome Project database at the Martinsried Institute for Protein of two nucleotides to yield the predicted protein Hxt12. The DNA
Sequences or the Saccharomyces Genome Database at Stanford sequence represents either a pseudogene or a sequencing error
University (except HXT4; see note d), and translated using GCG e Two models for the HXT14 open reading frame have been pro-

TRANSLATE (Genetics Computer Group 1994)
b Determined by the algorithm of Sharp and Li (1987) using the
Virtual Genome Center World Wide Web server at the University
of Minnesota
c The ORF in the HXT4 sequence of GenBank sequence U00060
has a deletion with respect to that of M81960 and X67321 result-
ing in a frameshift. The protein predicted by the latter two se-
quences is used in this study
posed (Maftahi et al. 1995). One (GenBank Z46259) invokes an
intron at the 5′ end of the gene, and the other (PIR2:S63299) does
not. The sequence of the protein predicted by the intronless open
reading frame is more consistent with the other members of the
Hxt family and is used in this study

HXT8 and HXT9 are near one another towards the end of
the left arm of chromosome X. The protein encoded by
HXT9, the telomere-proximal of these two, is 97% identi-
cal to the products of HXT11 and HXT12, which are linked
to the telomeres of chromosomes XV-L and IX-L, respec-
tively. Each of these genes is transcribed towards the
telomere. HXT13, HXT15, HXT16, and HXT17 are a quar-
tet of genes with 90–99% identity among their protein
products. Each is subtelomeric and transcribed in the di-
rection of the telomere.
The remainder of the HXT genes occupy internal posi-
tions on their respective chromosomes. Two tightly linked
triads of HXT genes occur: HXT1, HXT4 and HXT5 on the
right arm of chromosome VIII and HXT3, HXT6 and
HXT7 on the right arm of chromosome IV. The proteins
encoded by HXT6 and HXT7 are virtually identical and this
pair is highly similar to the HXT4 gene product (83.4%
identity). The product of HXT1 shares 86.4% identity with
the product of HXT3. In contrast, the product of HXT5 is
more similar to the low affinity glucose transporter of
Kluyveromyces lactis encoded by RAG1 than it is to any
of the S. cerevisiae HXT gene products (Fig. W1).
The large size of the HXT gene family is clearly due to
multiple gene duplications. Many of these duplications
have occurred by recombination among subtelomeric re-
gions. Genes encoding sugar transporters have been shared
among the telomeres of chromosomes IV, V, X and XIV,
and of chromosomes IX, X and XV. It is a common fea-
ture of many genes involved in carbon metabolism that
they occur as families distributed in subtelomeric posi-
tions in multiple chromosomes. Examples include the
SUC (Carlson et al. 1985), MAL (Michels et al. 1992), and
MEL (Turakainen et al. 1993) gene families. The HXT
family is unusual in the arrangement of its subtelomeric
members, in that they include cases of both linked and un-
linked genes. The family is also unusual among these car-
bon metabolism genes in that most of the family members
are not subtelomeric. The high identity among subtelom-
eric members of the family suggests that they are func-
tionally redundant or at least equivalent. They may be po-
sitioned too far from the ends of the chromosomes to be
subject to telomeric silencing (Renauld et al. 1993), but
their level of expression has not been determined.
Subtelomeric regions are populated with an unusually
high proportion of pseudogenes and fragmentary ORFs
(Bussey et al. 1995; Murakami et al. 1995); the HXT12 se-
quence includes a 2-bp insertion that would render it a
pseudogene. It has been suggested that this is a sequenc-

ing error (Nelissen et al. 1995), since the rest of the se-
quence is highly conserved with other HXT genes, but it
may instead be a recently formed pseudogene.
The high levels of recombination and gene duplication
at telomeres have been proposed to be a consequence of
mechanisms that maintain small chromosomes above a
critical minimal length required for stability (Bussey et al.
1995). However, the chromosomes that bear HXT genes
on their telomeres include the longest two chromosomes
(IV and XV) and do not include the shortest three chro-
mosomes (I, VI and III).
The HXT genes that are located internally in their chro-
mosomes may have arisen by tandem duplication, fol-
lowed by sequence divergence and relocation within the
genome by translocation or other means; the HXT6/HXT7
pair is obviously at an early stage of such a process. The
time of divergence among the other HXTs is not so clear.
The fact that the K. lactis RAG1 gene falls within the HXT
family (allied with HXT5; Fig. W1), and that RAG1 is one
of only two sugar transporter genes to have been found so
far in K. lactis (M. Wéselowski-Louvel, personal commu-
nication), suggests that most of the multitude of HXT
genes in S. cerevisiae have arisen quite recently in evolu-
tionary terms, perhaps as a consequence of artificial se-
lection on this yeast for performance in biotechnological
fermentations.
Expression and function
The codon adaptation index (Sharp and Li 1987) of the
hexose transporter genes ranges from 0.107 to 0.551
(Table 1). The codon adaptation index quantifies the syn-
onymous codon usage bias of a gene and is generally high
(up to 0.9) in highly expressed yeast genes (Sharp and
Lowe 1991). Hexose transporter genes with low indices
are thus predicted to be expressed at quite low levels,
while those with indices greater than 0.3 may be ex-
pressed at moderate levels.
The specific roles of the various hexose transporters
are not known. Hexose transport is required for the supply
of substrates to glycolysis, and as a source of carbon for
biosynthetic pathways, e.g., cell wall synthesis. Glucose
is involved in various signalling phenomena as well, and
hexose transporters may participate in these activities. It
is likely that the different transporters are suited for dif-
ferent functions and are expressed in response to different
environmental or cellular states. The large number of hex-
ose transporter genes in this unicellular microbe is re-
markable. In mammals, six hexose transporter genes and
one pseudogene are known with homology to the yeast
genes (GLUT1–GLUT7). These are differentially expressed
in different cells and tissues and have distinct roles on the
basis of their distribution, functional characteristics, and
cellular trafficking (Gould and Holman 1993).
Mutations in GAL2 drastically reduce growth on galac-
tose (Douglas and Condie 1954) and galactose transport
(Kuo et al. 1970), and GAL2 is only expressed in cells
growing in galactose (Sierkstra et al. 1992; Tschopp et al.
289
1986). These factors make it unique among the hexose
transporters in being able to support growth on galactose
as the sole carbon source.
Mutations in SNF3 are impaired in growth on low glu-
cose or raffinose (Neigeborn and Carlson 1984) and in
high-affinity glucose transport (Bisson et al. 1987). These
defects are suppressed by overexpression of HXT1, HXT2,
HXT3 and HXT4 (Bisson et al. 1987; Ko et al. 1993;
Kruckeberg and Bisson 1990; Lewis and Bisson 1991;
Theodoris et al. 1994) and by mutations in the RGT1 and
RGT2 genes (Marshall-Carlson et al. 1991). RGT1 may be
a transcriptional repressor of HXT genes (Özcan and
Johnston 1995). RGT2 is homologous to SNF3 (Fig. 1)
and the suppressing mutation is dominant (Marshall-Carl-
son et al. 1991; L. F. Bisson, personal communication).
SNF3 is expressed at a much lower level than the HXT
genes that have been analyzed (Özcan and Johnston 1995;
Kruckeberg, unpublished results; P. Vagnoli and L. F. Bis-
son, personal communication). It is probably not able to
contribute significantly to the flux of hexoses into the cell
and may not be a functional transporter. A number of ob-
servations suggest that it acts by regulating the expression
or activity of Hxt proteins. First, Hxt2 does not contribute
to high-affinity glucose transport when expressed nor-
mally in snf3 mutant cells (Wendell and Bisson 1994).
Second, a hxt1 hxt2 hxt3 hxt4 quadruple null mutant strain
is able to grow on low glucose, but only if a null allele of
SNF3 is present; wild-type SNF3 prevents growth of the
quadruple null strain on low glucose, presumably by neg-
atively regulating expression or function of another hex-
ose transporter gene(s) (Ko et al. 1993). Third, both high-
and low-affinity hexose transport kinetics are affected by
snf3 null mutations (Coons et al. 1995; Walsh et al. 1994).
None of the HXT genes examined is essential for via-
bility. Reifenberger et al. (1995) have constructed a strain
bearing null mutations in HXT1 through HXT7 (the hxt
null strain) that is unable to grow on glucose, fructose or
mannose and does not transport glucose at an appreciable
rate. This strain has proven useful for assessing the func-
tional properties of individual HXT genes.
Lewis and Bisson (1991) found that a null mutation of
HXT1 results in diminished high-affinity transport of glu-
cose and mannose. Reifenberger et al. (1995) found that
HXT1 restores relatively low affinity glucose transport to
the hxt null strain.
The HXT1 promoter is most active in early stages of a
batch fermentation (Ko et al. 1993; Lewis and Bisson
1991). These results are in agreement with the analysis of
HXT promoters by Özcan and Johnston (1995). They found
that the HXT1 promoter is only induced by high levels of
glucose. HXT1 expression is repressed in the absence of
glucose by the Rgt1 repressor. In the presence of glucose
this repression is relieved by the action of the Grr1 pro-
tein. A similar response to glucose is also seen for HXT2,
HXT3, and HXT4. However, only low concentrations of
glucose are stimulatory to HXT2 and HXT4 expression,
and HXT3 is induced in low or high glucose (Özcan and
Johnston 1995). As mentioned above, HXT1 is also induced
in high-osmolarity media; this regulation is exerted by the
290
Hog1 MAP kinase pathway (Hirayama et al. 1995). The
Hog1 pathway regulates a number of genes involved in
osmoregulation (Thevelein 1994). Perhaps the high glu-
cose concentrations required to stimulate HXT1 expression
do so via an osmotic effect as well as a glucose signal.
A null mutation in HXT2 impairs high-affinity trans-
port and growth on low glucose (Kruckeberg and Bisson
1990). When HXT2 is expressed in the hxt null strain of
Reifenberger et al. (1995), it confers high-affinity glucose
transport. HXT2 expression is induced when cells are
shifted to low-glucose media. In addition to regulation by
Rgt1, it is repressed in high glucose by the Mig1 repressor
protein. This dual control is responsive to the general glu-
cose-repression pathway, and involves the Hxk2, Reg1,
Snf1 and Ssn6 proteins (Özcan and Johnston 1995; Wen-
dell and Bisson 1994).
HXT3 confers only low-affinity transport on the hxt
null strain of Reifenberger et al. (1995). It is expressed in
both high and low glucose (Özcan and Johnston 1995),
but is maximally expressed at the onset of stationary
phase (Ko et al. 1993; Riou et al. 1995).
Less is known about the other HXT genes. HXT4 is ex-
pressed only in low glucose (Özcan and Johnston 1995)
and imparts moderately low-affinity glucose transport to
the hxt null strain (Reifenberger et al. 1995). HXT6 and
HXT7 display high-affinity kinetics when expressed in the
hxt null strain (Reifenberger et al. 1995). HXT13 is regu-
lated by the Hap2 transcriptional activator (B. Daignan-
Fornier, personal communication), suggesting that it is re-
pressed by glucose and expressed in respiring cells.
Studies are under way in a number of laboratories to
confirm and extend the observations sketched out above
and to examine the function and regulation of the other
hexose transporters identified in yeast. These studies will
be aided by the availability of the sequences of all of the
transporter genes and the knowledge of the relationships
among them. For example, strains can be produced that
bear null mutations specifically in genes with highly sim-
ilar proteins (e.g., HXT6 and HXT7). As the diverse func-
tions of the various sugar transporters are clarified, se-
quence analysis can be used to guide mutagenesis of the
genes to identify those residues that confer functional
specificity.
Conclusions
The picture that is emerging from both experimental work
and sequence analysis is that at any given stage, a yeast
cell is expressing an array of hexose transporters. The var-
ious transporter species will differ in their abundance and
in their intrinsic affinity for hexoses. Their kinetic charac-
teristics may also be modulated by post-translational
modifications or by interactions with one another or other
proteins. The kinetic characteristics of that cell will thus
be a summation of the kinetic properties of the trans-
porters being expressed. With 20 transporters to choose
from, the transport characteristics of such a cell can be
very complex indeed!
Acknowledgements I would like to acknowledge the accom-
plishment of all those who contributed to the determination of the
yeast genome sequence and the bioinformatic processing of the
data. I am grateful to Diana Chu and Frank Becker for critically
reading the manuscript. This work was supported by the Bur-
roughs Wellcome Fund.
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