CHAPTER 15 REGULATION OF GENE EXPRESSION AND DEVELOPMENT IN EUKARYOTES
428
i-'igttrc 15.11) Transcription factors required for the accu
rate initiation of transcription by RNA polymerase II ititicro
f *Most tissues are controlled by rraro-aaing proteins
The model shown provides the best fit to the data available
regarding the roles of transcription factors A, B, D, and E
Factor D is believed to interact with the TATA box as the first
step in the initiation process. Factor A is believed to act next,
forming a complex with factor D and DNA. Factor B is the
third factor to join the transcription-initiation complex After
the addition of factor B, the complex partially protects the
DMA sequence from -10 to +10 (transcription initiation at +
l>from nudease digestion. RNA polymerase II must be added
before factor E will bind After E is present, sequences from +
20 to +30 are protected from nuclease digestion, suggesting
thai factor E covers this region of the DNA. The complete
complex is then capable of initiating transcription accurately
from the + I site. (After S. Buratowski a al. Cell 56:
549-561. 1 9 8 9 )
signals (-J5 recognition sequence plus - 10 Pribnow box).
However, RNA polymerase II of eukarvotes, which
transcribes most of the protein-encoding nuclear genes.c::i.i./
initiate transcription accurately in intro without the addition
of >o:ir accessun pniteins :irtieiieiiiHnrHSfripliOHfmtors
The requirement lor these transcription factors (Fig. 15 19)
provides the potential for additional sites of regulation of
transcription. Whether these proteins interact with other
transcription factors in a regulator)' fashion ot whether
determined. In either case, ihey yield a slightly mote complex
picture of the initiation of transcription in eukarvotes.
Eukaryolic Transcription Units Are Monogenic
1the enhancer (left) and their relationship to the promoter
Trie sequence of one of the rwo 72 base pair repeats plus
MECHANISMS OF RECUATIOK OF TRANSCRIPTION IN HIGHER EUKARYOTES
429
At present, our knowledge about the mechanisms by which
gene expression is regulated in eukarvotes is expanding
rapidly. We know that different sets of genes ate transcribed
in different cell types in higher eukaryotes, and we know that
the different patterns of gene expression in different
encoded by regulatory genes that act in sequence
during differentiation. Clearly, regulatory mechanisms acting
at the level of transcription are important in cell
differentiation. However, the molecular details of these
regulatory mechanisms are still being worked out, and many
important questions about differentiation promise to
challenge geneticists for years to come.
In higher eukaryotes, it does seem very clear that
operons are mil important, if they exist at all. Although there
is evidence for operons or operonlike units in the lower
eukaryotes (e.g., fungi), operons appear to be rare or
nonexistent in higher eukaryotes. Most of the mRNAs of
higher eukaryotes characterized to date are tihnnnjcnic
Nucleosotnetree gap
(contain the coding sequence of one structural gene). In a few
cases, the primary transcripts are polygenic and are cleaved
to produce monogenic mRNAs
'AIIIKIUCCTS and Silencers Modulate i muse i
ipnon ill liiikaryolcs
Eukaryolic genes are regulated by promoter elements located
just upstream (5') from the transcription-initiation sites in a
manner quite similar to the regulation of prokaryotic genes
(Chapter 1-1) However, as in the Case of the Drosopljila Ubx
gene (see Fig 15.16), these eukaryolic promoters may be
very complex with binding sites lor many different regulatory
proteins. In addition to the nearby promoters, many
eukaryolic genes are also regulated bv more distant L I S acting
et'enten's culled engineers and silencers As the names suggest.
en/amccrs increase transcription sndsilenc-!'< ttnrense
transcription of the regulated genes. Since enhancers appear
I O be mure common and are much better understood, the
following discussion will focus on the propenies of enhancer 0/5145
elements Origin of replication
The basic features of enhancers thai distinguish them
from promoters are as follows
I Enhancers can act tnrr rcluuetly lar-ge distancesup to seierjl
thousand nucleotide pans from the reguljted "Core-ele
ment
aenetsl
2 Enixmcers are orientation independent[hev function Figure ty.20 Structure of the enhancer of simian virus 40
equally well in either orientation, normal or in vened (NV-+0). (a) Electron micrograph of an SV40 mimchromo some
(turned end-for-end). showing nucleosomes except in the region of the enhancer.
5 Enhancers are position independent<hev Fiinnion Within the cell, the enhancer region is probably Covered with
sequence-specific transcription factors, each bound at its
equally well whether located upstream (5') tVoin a gene,
specific recognition site, (b) Diagram of ihe structure ol the
downstream (3') from a gene, or present within an inuon of SV'40 minichromosome showing the location of the enhancer
a gene. The enhancer is about 220 nucleotide pairs in lengthcovering
the region from nucleotide position 100 to about position 320
(c) Diagram showing the components
Enhancers are relatively large elements, up to several
hundred nucleotidepairs ir. length. They sometimes contain
repealed sequences that have partial enhancer activity by
themselves Most enhancer elements function in a complete or
partially tissue-specific manner, that is, they frequently will
only enhance ihe transcription of genes in specific target
Late RNAs Ll 12 13 Alternating
Pu/Py
14 15 16 17
(n>r>ersenspi.ive)
tissues (those tissues where the gene-product is needed).

The most extensively studied enhancer is Lhat present

on the miriicrtromosorne of simian virus 40 (SV40), a virus
of monkeys that can be investigated in cell cultures. The
complete SV40 enhancer is about 220 nucleotide pairs in
length (Fig. l5-20XThis region of the SV40
minichromosome is not packaged into nucleosomes (Fig.
15-20c). Presumably, the SV40 enhancer is covered with
protein transcription factors that prevent it from becoming
wrapped into nucleosomes by the histones. The SV49
enhancer contains two 72-nucleotide-pair direct repeats,
and deletion of both repeats eliminates enhancer activity. If
one of the direct repeats is deleted, the enhancer is su'll func-
tional. Early experiments demonstrated that the SV40
enhancer could be moved to any other location on the SV40
minichromosome without loss.of activity. More-
xcACA-v;f*rccAAACc*TGCAicrc

Alternating
Pu/Py

Hanking nonrepeated enhancer DNA is shown at the

bottom Sequences that are sensitive to nucleases SI and
DNase! are identified at the bottom along with the
conserved core element of the enhancer The regions
labeled Tl. T2. and T3 are equivalent to the operator
regions of prokaryotic operons (see Chapter 14). They bind
the SV40 T-anrigen (a protein), which then represses
iranscripcioK. TheSpl bind mg sites are sequences to which
the mammalian transcrip tion factor Spl binds. (Reproduced
with permission from E Serfling, M. Jasin. and VP. Schaffner,
Trends in Genet. 1: 224-230. 1985 )