representation of a gene, transcription and translation. In this example, the gene comprises three exons with enhancer elements in the 5′ and 3′ flanking regions and a silencer element upstream of the promoter. UTR, untranslated region; Met, methionine (encoded by the start codon).
There is another layer of complexity governing how genes are expressed. Epigenetics is the study of how gene expression is regulated by mechanisms beyond the precise DNA sequence. Methylation of DNA around genes tends to silence expression. Modification to histones, such as acetylation or methylation, alters the chromatin structure to make enhancers and promoters either accessible or inaccessible to transcription factors. Acetylation tends to open up the chromatin structure, facilitating gene expression, whereas methylation tends to close it down and silence transcription. Genomic imprinting is an epigenetic phenomenon involving DNA methylation and modifications to histones such that gene expression varies according to which parent the particular chromosome came from.
RNA contains ribose sugar moieties rather than deoxyribose. RNA polymerase attaches ribonucleotides together to generate a single strand of mRNA that correlates to the DNA code of the gene, except that in place of thymidine, a very similar nucleoside, uridine, is incorporated. The initial mRNA strand (pre‐mRNA) is processed so that intronic gene regions are removed and only the exonic sequences are ‘spliced’ together. Not all exonic regions encode protein; stretches at either end constitute the 5′ and 3′ untranslated regions (UTRs) (Figure 2.2). Within the 3′ UTR, mRNA transcription is terminated by a specific motif, the polyadenylation signal, ∼20 base pairs upstream of where the mRNA gains a stretch of adenosine residues. This polyA tail provides stability as the mRNA is moved from the nucleus to the cytoplasm for translation into protein.
Translation into protein
mRNA is transported to the ribosomes, where protein synthesis occurs by ‘translation’ (Figure 2.3a). The ribosomes are attached to the outside of the endoplasmic reticulum (ER), leading to the description of ‘rough ER’. The ribosome is an RNA–protein complex that reads the mRNA sequence in groups of three nucleotides, called a codon. Each codon represents an amino acid. The start codon is the sequence of A–U–G nucleotides (corresponding to ATG in the genomic DNA) and specifies the amino acid methionine (Figure 2.2). From this point onwards, translation continues until a ‘stop’ codon is encountered (UAA, UGA or UAG).
By understanding these normal events of gene transcription and protein translation, it becomes possible to appreciate how mutations (sequence errors) in the genomic DNA lead to a miscoded, and consequently malfunctional, protein (Box 2.2). An entire gene may be missing (‘deleted’) or duplicated. An erroneous base pair in an enhancer or the promoter region may impair binding of a critical transcription factor and lessen a gene’s expression. A similar error in a coding exon might translate a different amino acid or even miscode a premature stop codon. Small deletions or insertions of one or two base pairs throw the whole triplet code out of frame. A mutation at the boundary between an intron and an exon can corrupt splicing so that the intron is included in the mature mRNA. All of these genetic events can affect endocrinology either as developmental disorders when that the fault is likely to be present in all or many of the body’s cells, or as acquired change later in life, potentially predisposing to the formation of an endocrine tumour (Chapter 10).
Figure 2.3 (a) Peptide hormone‐synthesizing and (b) steroid hormone‐synthesizing cells. In (b), cholesterol enters the cell via the low‐density lipoprotein receptor which is internalized.
Box 2.2 Genetic, genomic and epigenetic abnormalities that can result in endocrinopathy
Abnormalities in DNA (genetic)
Base substitution – swapping different nucleotides
Insertion or deletion – alters frame if exonic and not a multiple of three
Chromosomal abnormalities (genomic)
Numerical – loss of an X chromosome as in Turner syndrome (45,XO)
Structural:Inversion – region of a chromosome is turned upside downTranslocation – regions swapped between chromosomesDuplication – region of a chromosome is present twiceDeletion – region of a chromosome is excised and lost
Imprinting abnormalities (epigenetic)
Methylation – altered methylation changing local gene expression, such as Beckwith–Wiedemann syndrome with neonatal hypoglycaemia or transient neonatal diabetes mellitus associated with over‐expression of the gene called PLAGL1
Structural chromosomal abnormalities (above) can also cause imprinting errors
Post‐translational modification of peptides
Some polypeptides can function as hormones after little more than removal of the starting methionine, e.g. thyrotrophin‐releasing hormone (TRH), which comprises only three amino acids. Larger peptides can fold into three‐dimensional structures, which may contain helical or pleated domains. These shapes provide stability and affect how one protein interacts with another (e.g. how a hormone might bind to its receptor).
For hormones that require secretion out of the cell, additional modifications are important (Figure 2.4). The precursor peptide, called a pre‐prohormone, carries a lipophilic signal peptide at the amino terminus. This sequence is recognized by channel proteins so that the immature peptide can cross the ER membrane. Once inside the ER, the signal peptide is excised in preparation for other post‐translational changes (Figure 2.4a–d).
Figure 2.4 Potential post‐translational modifications of peptide hormones. Four types are shown. (a) Simple changes such as removal of the amino‐terminal ‘pro‐’ extension prior to secretion (e.g. parathyroid hormone) or the addition of intra‐chain disulphide bonds (e.g. growth hormone). (b) Multiple processing of a ‘polyprotein’ into a number of different
